14-hydroxy-docosahexaenoic acid compounds

ABSTRACT

The invention describes novel 14-hydroxy docosahexaenoic acid (DHA) analogs, their preparation, isolation, identification, purification and uses thereof.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a Section 371 National Stage Application ofInternational No. PCT/US2009/056998, filed Sep. 15, 2009 and publishedas WO 2010/033509 A2 on Mar. 25, 2010, which claims priority from U.S.Provisional Application 61/097,328, filed Sep. 16, 2008, and U.S.Provisional Application 61/138,652, filed Dec. 18, 2008, the contents ofwhich are incorporated herein in their entirety for all purposes.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH

The work leading to this invention was supported in part by NationalInstitutes of Health (NIH) grants P50-DE016191 and R37-GM038765. TheU.S. Government therefore may have certain rights in the invention.

FIELD OF THE INVENTION

The invention relates generally to novel dihydroxy analogues ofdocosahexaenoic acid (DHA) all having a hydroxyl group at C-14 of thecarbon chain and a second hydroxyl group at either the C-4, C-7 or C-13positions of the carbon chain.

BACKGROUND OF THE INVENTION

Given the contribution of uncontrolled inflammation to many humandiseases, the identification of endogenous control mechanisms in theacute inflammatory response is of wide interest (1). Classic lipidmediators such as the prostaglandins and leukotrienes are wellappreciated for their important pro-inflammatory roles in inflammation(2). In recent years, the resolution of inflammation has emerged as anarea with considerable potential to contain local mediators that may beuseful for new therapeutic approaches (for reviews, see 3, 4). Using anunbiased systems approach employing lipidomics, proteomics and celltrafficking to study self-resolving inflammatory exudates revealed thatthe termination of acute inflammation involves active biosyntheticprocesses producing novel endogenous lipid mediators that are bothanti-inflammatory and pro-resolving (5-8). It is now clear thatresolution of acute inflammation is an active rather than passiveprocess as previously understood (9), generating novel potentcounter-regulatory mediators termed resolvins and protectins (for arecent review, see Ref. 4).

Resolvins and protectins are biosynthesized by exudates from essentialomega-3 fatty acids (e.g., EPA and DHA), and the structures areestablished for key members of these families (4). The immune regulatoryactions of omega-3 fatty acids and their roles in human health anddiseases such as cancer and neuroinflammation are widely appreciated(10-12). Although omega-3 fatty acids are in wide use as dietarysupplements and potential therapeutics in many diseases includinginflammatory diseases, their mechanism(s) and connection to inflammationremain of interest. Resolvins and protectins display potent multi-levelanti-inflammatory and pro-resolving actions (13) and are members of anew genus of endogenous mediators of resolution (4). For example,resolvin E1 is biosynthesized from EPA and interacts with specificreceptors to control inflammatory cells (14, 15). Also, fat-1 transgenicmice, producing higher endogenous levels of omega-3, show reducedinflammatory status and elevated levels of resolvins and protectins,which when administered reduce inflammation and stimulate resolution(16-18). The main biosynthetic route with DHA for resolvins andprotectins proceeds during resolution via a17S-hydroperoxydocosahexaenoic intermediate produced by a lipoxygenasemechanism. With aspirin therapy, acetylated cyclooxygenase-2 producesaspirin-triggered 17R-epimers of resolvins and protectins as well asenhances their formation (6). Genetic deficiency or overexpression ofmurine 12/15-LOX regulates production of resolvins and protectins andalters their responses to both thermal injury and extent ofatherosclerosis (17, 18).

Therefore, a need exists for a further understanding of, an explorationor and identification of new useful materials previously not appreciatedas potent biological mediators of interest.

BRIEF SUMMARY OF THE INVENTION

Evidence for a new pathway of mediators operative in resolution of acuteinflammation that possess potent actions with PMN and MΦs is provided.Identification of these new mediators, coined maresins (macrophagemediators in resolving inflammation), provides evidence for autacoidsproduced from essential omega-3 fatty acids by a new pathway that may belinked to homeostasis, inflammation-resolution, wound healing andcancer.

Thus, the present invention provides evidence for new mediators and apathway operative during the resolution of acute inflammation thatconverts DHA to a novel 14S-series of DHA analogues that possess potentdual anti-inflammatory and pro-resolving actions with both neutrophilsand macrophages. Identification of these new 14S-series DHA analoguesprovides further evidence for local mediators produced from essentialomega-3 fatty acids that may link the known beneficial actions of DHA inorgan systems as well as actions reducing inflammatory disease andcancer in humans.

The present invention surprisingly provides novel compounds,compositions and methods of use pertaining to dihydroxy analogues ofdocosahexaenoic acid (DHA) all having a hydroxyl group at C-14 of thecarbon chain and a second hydroxyl group at either the C-4, C-7 or C-14positions of the carbon chain. These materials are biogenically derivedand isolated from media.

In one embodiment, the invention pertains to a new and useful DHAanalogue such as a compound comprising the formula (I):

wherein each of P₁ and P₂ individually is a protecting group or ahydrogen atom;

wherein

is a double bond;

wherein Z is —C(O)OR^(d), —C(O)NR^(c)R^(c), —C(O)H, —C(NH)NR^(c)R^(c),—C(S)H, —C(S)OR^(d), —C(S)NR^(c)R^(c), or —CN;

each R^(a), is independently selected from hydrogen, (C1-C6) alkyl,(C3-C8) cycloalkyl, cyclohexyl, (C4-C11)cycloalkylalkyl, (C5-C10)aryl,phenyl, (C6-C16) arylalkyl, benzyl, 2-6 membered heteroalkyl, 3-8membered cycloheteroalkyl, morpholinyl, piperazinyl, homopiperazinyl,piperidinyl, 4-11 membered cycloheteroalkylalkyl, 5-10 memberedheteroaryl or 6-16 membered heteroarylalkyl;

each R^(c), is independently a protecting group or R^(a), or,alternatively, each R^(c) is taken together with the nitrogen atom towhich it is bonded to form a 5 to 8-membered cycloheteroalkyl orheteroaryl which may optionally include one or more of the same ordifferent additional heteroatoms and which may optionally be substitutedwith one or more of the same or different R^(a) or suitable R^(b)groups;

each R^(b) is independently selected from ═O, —OR^(d),(C1-C3)haloalkyloxy, —OCF₃, ═S, —SR^(d), ═NR^(d), ═NOR^(d),—NR^(c)R^(c), halogen, —CF₃, —CN, —NC, —OCN, —SCN, —NO, —NO₂, ═N₂, —N₃,—S(O)R^(d), —S(O)₂R^(d), —S(O)₂OR^(d), —S(O)NR^(c)R^(c),—S(O)₂NR^(c)R^(c), —OS(O)R^(d), —OS(O)₂R^(d), —OS(O)₂OR^(d),—OS(O)₂NR^(c)R^(c), —C(O)R^(d), —C(O)OR^(d), —C(O)NR^(c)R^(c),—C(NH)NR^(c)R^(c), —C(NR^(a))NR^(c)R^(c), —C(NOH)R^(a),—C(NOH)NR^(c)R^(c), —OC(O)R^(d), —OC(O)OR^(d), —OC(O)NR^(c)R^(c),—OC(NH)NR^(c)R^(c), —OC(NR^(a))NR^(c)R^(c), —[NHC(O)]_(n)R^(d),—[NR^(a)C(O)]_(n)R^(d), —[NHC(O)]_(n)OR^(d), —[NR^(a)C(O)]_(n)R^(d),—[NHC(O)]_(n)NR^(c)R^(c), —[NR^(a)C(O)O]_(n)NR^(c)R^(c),—[NHC(NH)]_(n)NR^(c)R^(c) or —[NR^(a)C(NR^(a))]_(n)NR^(c)R^(c);

each n, independently is an integer from 0 to 3; and

each R^(d), independently is a protecting group or R^(a);

or a pharmaceutically acceptable salt thereof, provided when Z is—C(O)OR^(d), then R^(d) for Z is not a hydrogen. In certain aspects, P₁and P₂ are both hydrogen atoms. In another aspect, the double bonds atthe 4, 10, 16 and 19 positions are each of Z configuration or the doublebonds at the 4, 16 and 19 positions are each of the Z configuration.

A particular isomer of interest of the DHA analogue (I) is (Ia)comprising the formula:

Another isomer of interest of the DHA analogue (I) is (Ib) comprisingthe formula:

referred to as the “double dioxygenation” product.

It should be understood that compounds (Ia) and (Ib) include allpharmaceutically acceptable salts, esters thereof, the purified/isolatedforms, as well as compounds wherein one or both of the hydroxyls areconverted into a protecting group as described herein.

In another aspect, the present invention provides new and useful DHAanalogues such as a purified compound comprising the formula (I):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d) and n are as previously defined. In oneaspect, P₁ and P₂ are both hydrogen atoms. In another aspect, Z is—C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In another embodiment,the double bonds at the 4, 10, 16 and 19 positions are each of Zconfiguration or the double bonds at the 4, 16 and 19 positions are eachof the Z configuration. In still another embodiment, the 7 hydroxyl hasan S configuration. In still yet another embodiment, the 14 hydroxyl hasan S configuration.

In another aspect, the present invention provides new and useful DHAanalogues such as a purified compound comprising the formula (Ic):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d) and n are as previously defined. R₁ isselected from (C1-C6)alkyl, (C3-C8)cycloalkyl, cyclohexyl,(C4-C11)cycloalkylalkyl, (C5-C10)aryl, phenyl, (C6-C16)arylalkyl,benzyl, 2-6 membered heteroalkyl, 3-8 membered cycloheteroalkyl,morpholinyl, piperazinyl, homopiperazinyl, piperidinyl, 4-11 memberedcycloheteroalkylalkyl, 5-10 membered heteroaryl or 6-16 memberedheteroarylalkyl. In one aspect, R₁ is a methyl group.

In one aspect, P₁ and P₂ are both hydrogen atoms. In another aspect, Zis —C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In another embodiment,the double bonds at the 4, 10, 16 and 19 positions are each of Zconfiguration or the double bonds at the 4, 16 and 19 positions are eachof the Z configuration. In still another embodiment, the 7 hydroxyl hasan S configuration. In still yet another embodiment, the 14 hydroxyl hasan S configuration.

In another aspect, the present invention provides new and useful DHAanalogues such as a purified compound comprising the formula (Id):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d) and n are as previously defined. R₂ isselected from (C1-C6)alkyl, (C3-C8)cycloalkyl, cyclohexyl,(C4-C11)cycloalkylalkyl, (C5-C10), phenyl, (C6-C16)arylalkyl, benzyl,2-6 membered heteroalkyl, 3-8 membered cycloheteroalkyl, morpholinyl,piperazinyl, homopiperazinyl, piperidinyl, 4-11 memberedcycloheteroalkylalkyl, 5-10 membered heteroaryl or 6-16 memberedheteroarylalkyl. In one aspect, R₂ is a methyl group.

In one aspect, P₁ and P₂ are both hydrogen atoms. In another aspect, Zis —C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In another embodiment,the double bonds at the 4, 10, 16 and 19 positions are each of Zconfiguration or the double bonds at the 4, 16 and 19 positions are eachof the Z configuration. In still another embodiment, the 7 hydroxyl hasan S configuration. In still yet another embodiment, the 14 hydroxyl hasan S configuration.

In another aspect, the present invention provides new and useful DHAanalogues such as a purified compound comprising the formula (Ie):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d), R₁, R₂ and n are as previously defined.

In one aspect, P₁ and P₂ are both hydrogen atoms. In another aspect, R₁and R₂ are both methyl groups. In still another aspect, Z is —C(O)OR^(d)and R^(d) of Z is a hydrogen atom. In another embodiment, the doublebonds at the 4, 10, 16 and 19 positions are each of Z configuration orthe double bonds at the 4, 16 and 19 positions are each of the Zconfiguration. In still another embodiment, the 7 hydroxyl has an Sconfiguration. In still yet another embodiment, the 14 hydroxyl has an Sconfiguration.

In another aspect, the invention pertains to a new and useful DHAanalogue such as a compound comprising the formula (II):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d) and n are as previously defined,provided when Z is —C(O)OR^(d), then R^(d) for Z is not a hydrogen. Inone embodiment, P₁ and P₂ are both hydrogen atoms. In anotherembodiment, the double bonds at the 7, 16 and 19 positions are each of Zconfiguration.

In another aspect, the present invention provides new and useful DHAanalogues such as a purified compound comprising the formula (II):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d) and n are as previously defined. In oneaspect, P₁ and P₂ are both hydrogen atoms. In another aspect, Z is—C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In still another aspect,the double bonds at the 7, 16 and 19 positions are each of Zconfiguration.

In another aspect, the present invention provides new and useful DHAanalogues such as a compound comprising the formula (IIa):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d), R₁ and n are as previously defined. Inone aspect, P₁ and P₂ are both hydrogen atoms. In another aspect, Z is—C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In still another aspect,the double bonds at the 7, 16 and 19 positions are each of Zconfiguration. In yet another aspect, R₁ is a methyl group.

In another aspect, the present invention provides new and useful DHAanalogues such as a compound comprising the formula (IIb):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d), R₂ and n are as previously defined. Inone aspect, P₁ and P₂ are both hydrogen atoms. In another aspect, Z is—C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In still another aspect,the double bonds at the 7, 16 and 19 positions are each of Zconfiguration. In yet another aspect, R₂ is a methyl group.

In another aspect, the present invention provides new and useful DHAanalogues such as a compound comprising the formula (IIc):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d), R₁, R₂ and n are as previously defined.In one aspect, P₁ and P₂ are both hydrogen atoms. In another aspect, Zis —C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In still anotheraspect, the double bonds at the 7, 16 and 19 positions are each of Zconfiguration. In yet another aspect, R₁ and R₂ are both methyl groups.

In another aspect, the present invention provides new and useful DHAanalogues such as a compound comprising formula (III):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d) and n are as previously defined,provided when Z is —C(O)OR^(d), then R^(d) for Z is not a hydrogen. Inone embodiment, P₁ and P₂ are both hydrogen atoms. In anotherembodiment, the double bonds at the 4, 7, 16 and 19 positions are eachof Z configuration.

In still another aspect, the present invention provides new and usefulDHA analogues such as a purified compound comprising the formula (III):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d) and n are as previously defined. In oneaspect, P₁ and P₂ are both hydrogen atoms. In another aspect, Z is—C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In another embodiment,the double bonds at the 4, 7, 16 and 19 positions are each of Zconfiguration.

In still another aspect, the present invention provides new and usefulDHA analogues such as a compound comprising the formula (IIIa):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d), R₁ and n are as previously defined. Inone aspect, P₁ and P₂ are both hydrogen atoms. In another aspect, Z is—C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In still another aspect,R₁ is a methyl group. In another embodiment, the double bonds at the 4,7, 16 and 19 positions are each of Z configuration.

In still another aspect, the present invention provides new and usefulDHA analogues such as a compound comprising the formula (IIIb):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d), R₂ and n are as previously defined. Inone aspect, P₁ and P₂ are both hydrogen atoms. In another aspect, Z is—C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In still another aspect,R₂ is a methyl group. In another embodiment, the double bonds at the 4,7, 16 and 19 positions are each of Z configuration.

In still another aspect, the present invention provides new and usefulDHA analogues such as a compound comprising the formula (IIIc):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d), R₁, R₂ and n are as previously defined.In one aspect, P₁ and P₂ are both hydrogen atoms. In another aspect, Zis —C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In still anotheraspect, R₁ and R₂ are both methyl groups. In another embodiment, thedouble bonds at the 4, 7, 16 and 19 positions are each of Zconfiguration.

In yet another aspect, the present invention provides new and useful DHAanalogues such as a compound comprising formula (IV):

wherein

, Z, R^(a), R^(b), R^(c), R^(d) and n are as previously defined,provided when Z is —C(O)OR^(d), then R^(d) for Z is not a hydrogen. Inone embodiment, the double bonds at the 4, 7, 16 and 19 positions areeach of Z configuration.

In still another aspect, the present invention provides new and usefulDHA analogues such as a purified compound comprising the formula (IV):

wherein

, Z, R^(a), R^(b), R^(c), R^(d) and n are as previously defined. In anaspect, Z is —C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In anotherembodiment, the double bonds at the 4, 7, 16 and 19 positions are eachof Z configuration.

In another aspect, the invention provides a compound comprising theformula (V):

wherein each of P₁, P₂ and P₃ individually is a protecting group or ahydrogen atom and

, Z, R^(a), R^(b), R^(c), R^(d) and n are as previously defined,provided when Z is —C(O)OR^(d), then R^(d) for Z is not a hydrogen. Inone aspect, P₁, P₂ and P₃ are all hydrogen atoms. In another aspect, thedouble bonds at the 7, 16 and 19 positions are each of Z configuration.

In still yet another aspect, the present invention provides a purifiedcompound comprising the formula (V):

wherein P₁, P₂, P₃,

Z, R^(a), R^(b), R^(c), R^(d) and n are as previously defined. In oneaspect, P₁, P₂ and P₃ are all hydrogen atoms. In another aspect, Z is—C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In another embodiment,the double bonds at the 7, 16 and 19 positions are each of Zconfiguration.

In still yet another aspect, the present invention provides a compoundcomprising the formula (Va):

wherein P₁, P₂, P₃,

, Z, R^(a), R^(b), R^(c), R^(d), R₁ and n are as previously defined. Inone aspect, P₁, P₂ and P₃ are all hydrogen atoms. In another aspect, Zis —C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In another embodiment,the double bonds at the 7, 16 and 19 positions are each of Zconfiguration. In still another aspect, R₁ is a methyl group.

In another aspect, the present invention provides a compound comprisingthe formula (Vb):

wherein P₁, P₂, P₃,

, Z, R^(a), R^(b), R^(c), R^(d), R₂ and n are as previously defined. Inone aspect, P₁, P₂ and P₃ are all hydrogen atoms. In another aspect, Zis —C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In another embodiment,the double bonds at the 7, 16 and 19 positions are each of Zconfiguration. In still another aspect, R₂ is a methyl group.

In another aspect, the present invention provides a compound comprisingthe formula (Vc):

wherein P₁, P₂, P₃,

, Z, R^(a), R^(b), R^(c), R^(d), R₃ and n are as previously defined. Inone aspect, P₁, P₂ and P₃ are all hydrogen atoms. In another aspect, Zis —C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In another embodiment,the double bonds at the 7, 16 and 19 positions are each of Zconfiguration. In still another aspect, R₃ is a methyl group.

In another aspect, the present invention provides a compound comprisingthe formula (Vd):

wherein P₁, P₂, P₃,

, Z, R^(a), R^(b), R^(c), R^(d), R_(I), R₂ and n are as previouslydefined. In one aspect, P₁, P₂ and P₃ are all hydrogen atoms. In anotheraspect, Z is —C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In anotherembodiment, the double bonds at the 7, 16 and 19 positions are each of Zconfiguration. In still another aspect, R₁ and R₂ are methyl groups.

In another aspect, the present invention provides a compound comprisingthe formula (Ve):

wherein P₁, P₂, P₃,

, Z, R^(a), R^(b), R^(c), R^(d), R₁, R₃ and n are as previously defined.In one aspect, P₁, P₂ and P₃ are all hydrogen atoms. In another aspect,Z is —C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In anotherembodiment, the double bonds at the 7, 16 and 19 positions are each of Zconfiguration. In still another aspect, R₁ and R₃ are methyl groups.

In another aspect, the present invention provides a compound comprisingthe formula (Vf):

wherein P₁, P₂, P₃,

, Z, R^(a), R^(b), R^(c), R^(d), R₂, R₃ and n are as previously defined.In one aspect, P₁, P₂ and P₃ are all hydrogen atoms. In another aspect,Z is —C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In anotherembodiment, the double bonds at the 7, 16 and 19 positions are each of Zconfiguration. In still another aspect, R₂ and R₃ are methyl groups.

In another aspect, the present invention provides a compound comprisingthe formula (Vg):

wherein P₁, P₂, P₃,

, Z, R^(a), R^(b), R^(c), R^(d), R₁, R₂, R₃ and n are as previouslydefined. In one aspect, P₁, P₂ and P₃ are all hydrogen atoms. In anotheraspect, Z is —C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In anotherembodiment, the double bonds at the 7, 16 and 19 positions are each of Zconfiguration. In still another aspect, R₁, R₂ and R₃ are all methylgroups.

In another aspect, the present invention provides new and useful DHAanalogues such as a purified compound comprising the formula (VI):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d) and n are as previously defined.

In one aspect, P₁ and P₂ are both hydrogen atoms. In another aspect, Zis —C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In another embodiment,the double bonds at the 4, 10, 16 and 19 positions are each of Zconfiguration or the double bonds at the 4, 16 and 19 positions are eachof the Z configuration. In still another embodiment, the 7 hydroxyl hasan S configuration. In still yet another embodiment, the 14 hydroxyl hasan S configuration.

In another aspect, the present invention provides new and useful DHAanalogues such as a compound comprising the formula (VIa):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d), R₁ and n are as previously defined. Inone aspect, R₁ is a methyl group.

In one aspect, P₁ and P₂ are both hydrogen atoms. In another aspect, Zis —C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In another embodiment,the double bonds at the 4, 10, 16 and 19 positions are each of Zconfiguration or the double bonds at the 4, 16 and 19 positions are eachof the Z configuration. In still another embodiment, the 7 hydroxyl hasan S configuration. In still yet another embodiment, the 14 hydroxyl hasan S configuration.

In another aspect, the present invention provides new and useful DHAanalogues such as compound comprising the formula (VIb):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d), R₂ and n are as previously defined. Inone aspect, R₂ is a methyl group.

In one aspect, P₁ and P₂ are both hydrogen atoms. In another aspect, Zis —C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In another embodiment,the double bonds at the 4, 10, 16 and 19 positions are each of Zconfiguration or the double bonds at the 4, 16 and 19 positions are eachof the Z configuration. In still another embodiment, the 7 hydroxyl hasan S configuration. In still yet another embodiment, the 14 hydroxyl hasan S configuration.

In another aspect, the present invention provides new and useful DHAanalogues such as a compound comprising the formula (VIc):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d), R₁, R₂ and n are as previously defined.

In one aspect, P₁ and P₂ are both hydrogen atoms. In another aspect, R₁and R₂ are both methyl groups. In still another aspect, Z is —C(O)OR^(d)and R^(d) of Z is a hydrogen atom. In another embodiment, the doublebonds at the 4, 10, 16 and 19 positions are each of Z configuration orthe double bonds at the 4, 16 and 19 positions are each of the Zconfiguration. In still another embodiment, the 7 hydroxyl has an Sconfiguration. In still yet another embodiment, the 14 hydroxyl has an Sconfiguration.

In another aspect, the C-14 alcohol has an S configuration for thecompounds noted throughout the application.

In another aspect, the present invention provides pharmaceuticalcompositions comprising one or more compounds of the invention, with orwithout other active pharmaceutical ingredients, in admixture with apharmaceutically acceptable vehicle. Such a preparation can beadministered according to the methods of the current invention.

In yet another aspect, the present invention is drawn to methods fortreating or preventing inflammation or inflammatory disease in a mammal.The method involves administering a prophylactically or therapeuticallyeffective amount of at least one compound of the invention, or apharmaceutical composition thereof. For example, the compounds of theinvention can be used to treat or prevent inflammation, cancer,neurodegeneration, memory loss, wrinkles, psoriasis, dandruff ordermatitis by administering to an individual in need thereof, aneffective amount of any of the compounds described herein.

Additionally, the compounds of the invention can be used to neuraldevelopment, fetal development, homeostasis, tissue remodeling, or woundrepair by administering to an individual in need thereof, an effectiveamount of any of the compounds described herein.

Additional features and advantages of the invention will become moreapparent from the following detailed description and claims.

While multiple embodiments are disclosed, still other embodiments of thepresent invention will become apparent to those skilled in the art fromthe following detailed description. As will be apparent, the inventionis capable of modifications in various obvious aspects, all withoutdeparting from the spirit and scope of the present invention.Accordingly, the detailed descriptions are to be regarded asillustrative in nature and not restrictive.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1. Self-resolving acute inflammatory exudates. FIG. 1A. Time courseof PMN (dotted line) accumulation, resolution and HDHA formation duringzymosan-initiated peritonitis. Exudates were extracted for targetedlipidomics using LC/MS/MS. Hydroxydocosahexaenoic acids, 17-HDHA (dashedline) and 14S-HDHA (solid) identified using MRM results arerepresentative (n=3) and PMN n=4. FIGS. 1B and 1C. Representative massspectra 17-HDHA (Panel B) and 14S-HDHA (Panel C), n=3.

FIG. 2. Macrophages generate novel products. FIG. 2A. Murine residentMΦs (5×10⁶ cells/ml) incubated with DHA or 14S-HpDHA: targetedLC/MS/MS-based mediator lipidomics. Selected ion chromatogram (m/z359/250) of 7,14-dihydroxydocosahexaenoic acid (II) and its transconjugated isomer (I). Selected ion chromatogram (dashed overlay; m/z359/250) shows double dioxygenation product 7S,14S-diHDHA. Inset: FACSof isolated resident MΦs. FIG. 2 Panel B. Lipid mediator lipidomics.Mass spectra for 7,14-dihydroxydocosahexaenoic acid (m/z 359) and (PanelC) corresponding isomer. See inset and text for diagnostic ions n=3.

FIG. 3. Anti-inflammatory novel macrophage products. FIG. 3A. Reductionin PMN murine peritonitis. Activity in methyl formate fractions from C18extraction of isolated MΦs (black), MΦ product (20 ng/mouse) isolatedwith RP-HPLC, PD1 (20 ng/mouse), or RvE1 (20 ng/mouse). Results areexpressed as exudate PMN mean±SEM (n=3, *, p<0.05, compared to zymosanplus vehicle). FIG. 3B Differential PMN vs. monocyte actions. Mice wereinjected with the double dioxygenation product (0.1 ng/mouse), MΦisolate (0.1 ng/mouse), or vehicle alone (as in Panel A), followed byi.p injection of zymosan (1 mg) to evoke peritonitis. After 2 h,leukocytes were enumerated. Black bar, PMN; hatched bar, mononuclearcells. Results are mean±SEM (n=3, *, p<0.05, compared to zymosan plusvehicle; †, p<0.05, double dioxygenation versus MΦ isolate). FIG. 3C.Reduction in peritonitis: Dose response. MΦ product isolated followingHPLC isolation was injected i.v.˜2 min before i.p. zymosan. Results aremean±SEM (n=3, *, p<0.05, compared to zymosan plus vehicle).

FIG. 3D. MaR1 enhances phagocytosis. MΦs (24-well plate, 10⁵ cells/well)were exposed to indicated concentrations for 15 min followed byFITC-labeled zymosan (30 min, 37° C.). Results are mean±SEM expressed as% increase above vehicle (n=3, *, p<0.05 compared to vehicle; †, p<0.05,double dioxygenation versus MaR1). Solid diamond, MaR1. Solid box,double dioxygenation product 7S,14S-diHDHA. Inset, Comparison of MaR1with other mediators [1 nM].

FIG. 4. Identification of methoxy trapping product from MΦs. MS/MSspectrum of m/z 373 product at 10.2 min. Inset: Extracted ionchromatogram of m/z 373-263 and deduced structure.

FIG. 5. Biosynthetic scheme proposed for maresin 1 and related products.Stereochemistries and double bond geometries of the new di-hydroxycontaining mediators are tentative assignments and depicted in likelyconfigurations based on biogenic synthesis, trapping and labeling.

FIG. 6 shows the MS-MS spectrum of the novel product obtained with H₂¹⁸O and MΦ incubations.

FIG. 7 is the GC-MS spectrum obtained for the 13,14-dihydroxy vicinaldiol from DHA and MΦs. This product derivative was obtained aftertreatment with diazomethane and BSTFA to give the methyl ester, OTMSderivative.

FIG. 8 is a general synthetic scheme to prepare one type of analog thatis applicable to di and tri hydroxy analogs.

FIG. 9 is a general synthetic scheme to prepare a terminal fluorinatedanalog.

FIG. 10 depicts Structures, LC-MS and GC-MS fragmentation for novel14-series compounds identified using mediator-based lipidomics.^(a)LC-MS/MS analysis was performed with an Agilent 1100 series HPLCcoupled to an ABI Sciex Instruments 3200 Qtrap linear ion trapquadrupole mass spectrometer equipped with an Agilent Eclipse Plus C18column (4.6 mm×50 mm×1.8 μm). The mobile phase consisted ofmethanol/water/acetic acid (60/40/0.01; v/v/v) and was ramped to80/20/0.1 (v/v/v) over 7.5 minutes and to 95/5/0.01 (v/v/v) over thenext 4.5 minutes at a flow rate of 400 μl/min. The flow rate wasdecreased to 200 μl/min for 3 minutes, then returned to 400 μl/min andthe mobile phase was ramped up over the next 6 minutes to 100/0/0.01(v/v/v) before returning to 60/40/0.01 (v/v/v). ^(b)GC-MS analysis wasperformed with an Agilent HP6890 equipped with a HP5973N mass detector.A HP-5MS column (30 m×0.25 mm×0.25 μm) was employed with a temperatureprogram; the initial temperature was 150° C., followed by 230° C. (8min) and 280° C. (10 min) with a helium flow rate of 1.0 ml/min.Trimethylsilyl derivatives were prepared following treatment withdiazomethane. Spectra were recorded in methanol using an Agilent 4682UV-Vis spectrophotometer or an Agilent 1100 series DAD.*Stereochemistries shown are tentative assignments. Double bondgeometries are shown in likely configurations based on proposedbiosynthetic pathways.

DETAILED DESCRIPTION

The endogenous cellular and molecular mechanisms that control acuteinflammation and its resolution are of wide interest. Usingself-resolving inflammatory exudates and lipidomics, a new pathwayinvolving biosynthesis of potent anti-inflammatory and pro-resolvingmediators from essential fatty acid docosahexaenoic acid (DHA) bymacrophages was identified. During the resolution of murine peritonitis,exudates accumulated both 17-HDHA, a known marker of 17S-D-seriesresolvin and protectin biosynthesis, and 14S-HDHA from endogenous DHA.Addition of either DHA or14S-hydroperoxydocosa-4Z,7Z,10Z,12E,16Z,19Z-hexaenoic acid (14S-HpDHA)to activated macrophages converted these substrates to noveldihydroxy-containing products that possessed potent anti-inflammatoryand pro-resolving activity with a potency similar to resolvin E1 andprotectin D1. Stable isotope incorporation, intermediate trapping, andcharacterization of physical and biological properties of the productsdemonstrated a novel 14-lipoxygenase pathway, generating bioactive7,14-dihydroxy-docosa-4Z,8,10,12,16Z,19Z-hexaenoic acid, coined maresin(macrophage mediator in resolving inflammation: MaR), which enhancesresolution. These findings provide that maresins and this new metabolomeare involved in some of the beneficial actions of DHA and macrophages intissue homeostasis, inflammation-resolution, wound healing and hostdefense.

Abbreviations used throughout the specification:

7S,14S-diHDHA (double dioxygenation),7S,14S-dihydroxydocosa-4Z,8E,10Z,12E,16Z,19Z-hexaenoic acid

14S-HDHA, 14S-hydroxydocosa-4Z,7Z,10Z,12E,16Z,19Z-hexaenoic acid

14S-HpDHA, 14S-hydroperoxydocosa-4Z,7Z,10Z,12E,16Z,19Z-hexaenoic acid

17S-HDHA, 17S-hydroxydocosa-4Z,7Z,10Z,13Z,15E,19Z-hexaenoic acid

DHA, docosahexaenoic acid

GC-MS, gas chromatography-mass spectrometry

LC/MS/MS, liquid chromatography-tandem mass spectrometry

LOX, lipoxygenase

MaR, Maresin, macrophage mediator in resolving inflammation

MΦ, macrophage

PD1, Protectin D1,10R,17S-dihydroxydocosa-4Z,7Z,11E,13E,15Z,19Z-hexaenoic acid

PGE₂, prostaglandin E₂

PMN, polymorphonuclear neutrophils

Rv, resolvin

RvD1, Resolvin D1, 7S,8R,17Strihydroxydocosa-4Z,9E,11E,13Z,15E,19Z-hexaenoic acid

RvE1, Resolvin E1,5S,12R,18R-trihydroxyeicosa-6Z,8E,10E,14Z,16E-pentaenoic acid

In the specification and in the claims, the terms “including” and“comprising” are open-ended terms and should be interpreted to mean“including, but not limited to . . . .” These terms encompass the morerestrictive terms “consisting essentially of” and “consisting of.”

It must be noted that as used herein and in the appended claims, thesingular forms “a”, “an”, and “the” include plural reference unless thecontext clearly dictates otherwise. As well, the terms “a” (or “an”),“one or more” and “at least one” can be used interchangeably herein. Itis also to be noted that the terms “comprising”, “including”,“characterized by” and “having” can be used interchangeably.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meanings as commonly understood by one of ordinary skillin the art to which this invention belongs. All publications and patentsspecifically mentioned herein are incorporated by reference in theirentirety for all purposes including describing and disclosing thechemicals, instruments, statistical analyses and methodologies which arereported in the publications which might be used in connection with theinvention. All references cited in this specification are to be taken asindicative of the level of skill in the art. Nothing herein is to beconstrued as an admission that the invention is not entitled to antedatesuch disclosure by virtue of prior invention.

“Compounds of the invention” refers to the di-hydroxy, trihydroxy,and/or epoxide DHA analogues and compounds encompassed by genericformulae disclosed herein and includes any specific compounds withinthose formulae whose structure is disclosed herein. The compounds of theinvention may be identified either by their chemical structure and/orchemical name. When the chemical structure and chemical name conflict,the chemical structure is determinative of the identity of the compound.The compounds of the invention may contain one or more chiral centersand/or double bonds and therefore, may exist as stereoisomers, such asdouble-bond isomers (i.e., geometric isomers), enantiomers ordiastereomers. Accordingly, the chemical structures depicted hereinencompass all possible enantiomers and stereoisomers of the illustratedcompounds including the stereoisomerically pure form (e.g.,geometrically pure, enantiomerically pure or diastereomerically pure)and enantiomeric and stereoisomeric mixtures. Enantiomeric andstereoisomeric mixtures can be resolved into their component enantiomersor stereoisomers using separation techniques or chiral synthesistechniques well known to the skilled artisan. The compounds of theinvention also include isotopically labeled compounds where one or moreatoms have an atomic mass different from the atomic mass conventionallyfound in nature.

The compounds depicted throughout the specification containethylenically unsaturated sites. Where carbon carbon double bonds exist,the configurational chemistry can be either cis (E) or trans (Z) and thedepictions throughout the specification are not meant to be limiting.The depictions are, in general, presented based upon the configurationalchemistry of related DHA or EPA compounds, and although not to belimited by theory, are believed to possess similar configurationchemistry. The use of

reflects this throughout the specification and claims so that both cisand trans isomers are contemplated. In certain embodiments theconfiguration of the ethylenic bond is known and is particularlydescribed.

In one aspect of the invention, the compound(s) of the invention aresubstantially purified and/or isolated by techniques known in the art.The purity of the purified compounds is generally at least about 90%,preferably at least about 95%, and most preferably at least about 99% byweight.

Thus, the term “purified” as used herein does not require absolutepurity; rather, it is intended as a relative term. For example, apurified DHA analogue can be one in which the subject DHA analogue is ata higher concentration than the analogue would be in its naturalenvironment within an organism. For example, a DHA analogue of theinvention can be considered purified if the analogue content in thepreparation represents at least 50%, 60%, 70%, 80%, 85%, 90%, 92%, 95%,98%, or 99% of the total analogue content of the preparation.

“Biological activity” and its contextual equivalents “activity” and“bioactivity” means that a compound elicits a statistically valid effectin any one biological test assays. Preferably, the threshold fordefining an “active” compound will be reproducible and statisticallyvalid effects of at least 25% deviation from untreated control atconcentrations at or lower than 1 μM.

“Biological test assay” means a specific experimental procedure.Non-limiting examples of biological test assays include: 1) ligandbinding, either direct or indirect, to a purified target, subcellularfraction, intact cell, or cell or tissue extract; 2) metabolicprotection with enhanced half-life when exposed to a purified target,subcellular fraction, intact cell, cell or tissue extract, oradministered to intact organism by any route; 3) prevention, reversal,or amelioration of cell- and tissue-based functional responsesrecognized by skilled artisans to represent surrogates foranti-inflammatory action (e.g., altered cytokine production andrelease); and 4) prevention, reversal, or amelioration of symptomsand/or disease processes in animal models of inflammation andinflammatory disease.

“Detectable label” means any chemical or biological modality which canbe used to track, trace, localize, quantify, immobilize, purify, oridentify compounds through appropriate means of detection known in theart. Non-limiting examples of detectable labels include fluorescence,phosphorescence, luminescence, radioactive or biospecific affinitycapture labels.

“Electronegative group” is a chemical group that tends to acquire ratherthan lose electrons in its chemical interactions. Examples ofelectronegative groups include, but are not limited to, —NO₂, ammoniumsalts, sulfonyl groups, carbonyl groups, halogens, esters, carboxylicacids, nitriles, etc.

“In Situ” refers to and includes the terms “in vivo,” “ex vivo” and “invitro” as these terms are commonly recognized and understood by theskilled artisan. Moreover, the phrase “in situ” is employed herein inits broadest connotative and denotative context to identify an entity,cell, or tissue as found or in place, without regard to its source ororigin, its condition or status or its duration or longevity at thatlocation or position.

“Pharmaceutically acceptable” means approved by a regulatory agency ofthe Federal or a state government or listed in the U.S. Pharmacopoeia orother generally recognized pharmacopoeia for use in animals, and moreparticularly in humans.

“Pharmaceutically acceptable salt” refers to a salt of a compound of theinvention that is pharmaceutically acceptable and that possesses thedesired pharmacological activity of the parent compound. Such saltsinclude: (1) salts fanned when an basic proton is present in the parentcompound such as acid addition salts, formed with inorganic acids suchas hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,phosphoric acid, and the like; or those formed with organic acids suchas acetic acid, propionic acid, hexanoic acid, cyclopentanepropionicacid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinicacid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid,benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelicacid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonicacid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid,4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid,4-toluenesulfonic acid, camphorsulfonic acid,4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid,3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid,lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoicacid, salicylic acid, stearic acid, muconic acid and the like; or (2)salts formed when an acidic proton is present in the parent compound andeither is replaced by a metal ion, e.g., an alkali metal ion, analkaline earth ion, or an aluminum ion; or coordinates with an organicbase such as ethanolamine, diethanolamine, triethanolamine,N-methylglucamine, triethylamine, propylamino, diazabicycloundecane andthe like.

“Pharmaceutically acceptable vehicle” refers to a diluent, adjuvant,excipient or carrier with which a compound of the invention isadministered.

The phrase “pharmaceutically acceptable carrier” as used herein means apharmaceutically acceptable material, composition or vehicle, such as aliquid or solid filler, diluent, excipient, solvent or encapsulatingmaterial, involved in carrying or transporting a compound(s) of thepresent invention within or to the subject such that it can perform itsintended function. Typically, such compounds are carried or transportedfrom one organ, or portion of the body, to another organ, or portion ofthe body. Each carrier must be “acceptable” in the sense of beingcompatible with the other ingredients of the formulation and notinjurious to the patient. Some examples of materials which can serve aspharmaceutically acceptable carriers include: sugars, such as lactose,glucose and sucrose; starches, such as corn starch and potato starch;cellulose, and its derivatives, such as sodium carboxymethyl cellulose,ethyl cellulose and cellulose acetate; powdered tragacanth; malt;gelatin; talc; excipients, such as cocoa butter and suppository waxes;oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil,olive oil, corn oil and soybean oil; glycols, such as propylene glycol;polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol;esters, such as ethyl oleate and ethyl laurate; agar; buffering agents,such as magnesium hydroxide and aluminum hydroxide; alginic acid;pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol;phosphate buffer solutions; and other non-toxic compatible substancesemployed in pharmaceutical formulations.

“Prodrug” refers to a derivative of a drug molecule that requires atransformation within the body to release the active drug. Prodrugs arefrequently (though not necessarily) pharmacologically inactive untilconverted to the parent drug. A hydroxyl containing drug may beconverted to, for example, to a sulfonate, ester or carbonate prodrug,which may be hydrolyzed in vivo to provide the hydroxyl compound. Anamino containing drug may be converted, for example, to a carbamate,amide, imine, phosphonyl, phosphoryl or sulfenyl prodrug, which may behydrolyzed in vivo to provide the amino compound. A carboxylic acid drugmay be converted to an ester (including silyl esters and thioesters),amide or hydrazide prodrug, which be hydrolyzed in vivo to provide thecarboxylic acid compound. Prodrugs for drugs which contain differentfunctional groups other than those listed above are well known to theskilled artisan.

“Promoiety” refers to a form of protecting group that when used to maska functional group within a drug molecule converts the drug into aprodrug. Typically, the promoiety will be attached to the drug viabond(s) that are cleaved by enzymatic or non-enzymatic means in vivo.

“Protecting group” refers to a grouping of atoms that when attached to areactive functional group in a molecule masks, reduces or preventsreactivity of the functional group. Examples of protecting groups can befound in Green et al., “Protective Groups in Organic Chemistry”, (Wiley,2.sup.nd ed. 1991) and Harrison et al., “Compendium of Synthetic OrganicMethods,” Vols. 1-8 (John Wiley and Sons, 1971-1996). Representativeamino protecting groups include, but are not limited to, formyl, acetyl,trifluoroacetyl, benzyl, benzyloxycarbonyl (“CBZ”), tert-butoxycarbonyl(“Boc”), trimethylsilyl (“TMS”), 2-trimethylsilyl-ethanesulfonyl(“SES”), trityl and substituted trityl groups, allyloxycarbonyl,9-fluorenylmethyloxycarbonyl (“FMOC”), nitro-veratryloxycarbonyl(“NVOC”) and the like. Representative hydroxy protecting groups include,but are not limited to, those where the hydroxy group is either acylated(e.g., methyl and ethyl esters, acetate or propionate groups or glycolesters) or alkylated such as benzyl, and trityl ethers as well as alkylethers, tetrahydropyranyl ethers, trialkylsilyl ethers (e.g., TMS orTIPPS groups) and allyl ethers.

“Subject” means living organisms susceptible to conditions or diseasescaused or contributed to by inflammation, inflammatory responses,vasoconstriction and myeloid suppression. Examples of subjects includehumans, dogs, cats, cows, goats and mice. The term subject is furtherintended to include transgenic species such as, for example, transgenicmice.

“Alkyl” by itself or as part of another substituent refers to asaturated or unsaturated branched, straight-chain or cyclic monovalenthydrocarbon radical having the stated number of carbon atoms (i.e.,C1-C6 means one to six carbon atoms) that is derived by the removal ofone hydrogen atom from a single carbon atom of a parent alkane, alkeneor alkyne. Typical alkyl groups include, but are not limited to, methyl;ethyls such as ethanyl, ethenyl, ethynyl; propyls such as propan-1-yl,propan-2-yl, cyclopropan-1-yl, prop-1-en-1-yl, prop-1-en-2-yl,prop-2-en-1-yl, cycloprop-1-en-1-yl; cycloprop-2-en-1-yl,prop-1-yn-1-yl, prop-2-yn-1-yl, etc.; butyls such as butan-1-yl,butan-2-yl, 2-methyl-propan-1-yl, 2-methyl-propan-2-yl, cyclobutan-1-yl,but-1-en-1-yl, but-1-en-2-yl, 2-methyl-prop-1-en-1-yl, but-2-en-1-yl,but-2-en-2-yl, buta-1,3-dien-1-yl, buta-1,3-dien-2-yl,cyclobut-1-en-1-yl, cyclobut-1-en-3-yl, cyclobuta-1,3-dien-1-yl,but-1-yn-1-yl, but-1-yn-3-yl, but-3-yn-1-yl, etc.; and the like. Wherespecific levels of saturation are intended, the nomenclature “alkanyl,”“alkenyl” and/or “alkynyl” is used, as defined below. In preferredembodiments, the alkyl groups are (C1-C6)alkyl.

“Alkanyl” by itself or as part of another substituent refers to asaturated branched, straight-chain or cyclic alkyl derived by theremoval of one hydrogen atom from a single carbon atom of a parentalkane. Typical alkanyl groups include, but are not limited to,methanyl; ethanyl; propanyls such as propan-1-yl, propan-2-yl(isopropyl), cyclopropan-1-yl, etc.; butanyls such as butan-1-yl,butan-2-yl (sec-butyl), 2-methyl-propan-1-yl (isobutyl),2-methyl-propan-2-yl (t-butyl), cyclobutan-1-yl, etc.; and the like. Inpreferred embodiments, the alkanyl groups are (C1-C6)alkanyl.

“Alkenyl” by itself or as part of another substituent refers to anunsaturated branched, straight-chain or cyclic alkyl having at least onecarbon-carbon double bond derived by the removal of one hydrogen atomfrom a single carbon atom of a parent alkene. The group may be in eitherthe cis or trans conformation about the double bond(s). Typical alkenylgroups include, but are not limited to, ethenyl; propenyls such asprop-1-en-1-yl, prop-1-en-2-yl, prop-2-en-1-yl, prop-2-en-2-yl,cycloprop-1-en-1-yl; cycloprop-2-en-1-yl; butenyls such asbut-1-en-1-yl, but-1-en-2-yl, 2-methyl-prop-1-en-1-yl, but-2-en-1-yl,but-2-en-2-yl, buta-1,3-dien-1-yl, buta-1,3-dien-2-yl,cyclobut-1-en-1-yl, cyclobut-1-en-3-yl, cyclobuta-1,3-dien-1-yl, etc.;and the like. In preferred embodiments, the alkenyl group is(C2-C6)alkenyl.

“Alkynyl” by itself or as part of another substituent refers to anunsaturated branched, straight-chain or cyclic alkyl having at least onecarbon-carbon triple bond derived by the removal of one hydrogen atomfrom a single carbon atom of a parent alkyne. Typical alkynyl groupsinclude, but are not limited to, ethynyl; propynyls such asprop-1-yn-1-yl, prop-2-yn-1-yl, etc.; butynyls such as but-1-yn-1-yl,but-1-yn-3-yl, but-3-yn-1-yl, etc.; and the like. In preferredembodiments, the alkynyl group is (C2-C6)alkynyl.

“Alkyldiyl” by itself or as part of another substituent refers to asaturated or unsaturated, branched, straight-chain or cyclic divalenthydrocarbon group having the stated number of carbon atoms (i.e., C1-C6means from one to six carbon atoms) derived by the removal of onehydrogen atom from each of two different carbon atoms of a parentalkane, alkene or alkyne, or by the removal of two hydrogen atoms from asingle carbon atom of a parent alkane, alkene or alkyne. The twomonovalent radical centers or each valency of the divalent radicalcenter can form bonds with the same or different atoms. Typicalalkyldiyl groups include, but are not limited to, methandiyl; ethyldiylssuch as ethan-1,1-diyl, ethan-1,2-diyl, ethen-1,1-diyl, ethen-1,2-diyl;propyldiyls such as propan-1,1-diyl, propan-1,2-diyl, propan-2,2-diyl,propan-1,3-diyl, cyclopropan-1,1-diyl, cyclopropan-1,2-diyl,prop-1-en-1,1-diyl, prop-1-en-1,2-diyl, prop-2-en-1,2-diyl,prop-1-en-1,3-diyl, cycloprop-1-en-1,2-diyl, cycloprop-2-en-1,2-diyl,cycloprop-2-en-1,1-diyl, prop-1-yn-1,3-diyl, etc.; butyldiyls such as,butan-1,1-diyl, butan-1,2-diyl, butan-1,3-diyl, butan-1,4-diyl,butan-2,2-diyl, 2-methyl-propan-1,1-diyl, 2-methyl-propan-1,2-diyl,cyclobutan-1,1-diyl; cyclobutan-1,2-diyl, cyclobutan-1,3-diyl,but-1-en-1,1-diyl, but-1-en-1,2-diyl, but-1-en-1,3-diyl,but-1-en-1,4-diyl, 2-methyl-prop-1-en-1,1-diyl,2-methanylidene-propan-1,1-diyl, buta-1,3-dien-1,1-diyl,buta-1,3-dien-1,2-diyl, buta-1,3-dien-1,3-diyl, buta-1,3-dien-1,4-diyl,cyclobut-1-en-1,2-diyl, cyclobut-1-en-1,3-diyl, cyclobut-2-en-1,2-diyl,cyclobuta-1,3-dien-1,2-diyl, cyclobuta-1,3-dien-1,3-diyl,but-1-yn-1,3-diyl, but-1-yn-1,4-diyl, buta-1,3-diyn-1,4-diyl, etc.; andthe like. Where specific levels of saturation are intended, thenomenclature alkanyldiyl, alkenyldiyl and/or alkynyldiyl is used. Whereit is specifically intended that the two valencies are on the samecarbon atom, the nomenclature “alkylidene” is used. In preferredembodiments, the alkyldiyl group is (C1-C6)alkyldiyl. Also preferred aresaturated acyclic alkanyldiyl groups in which the radical centers are atthe terminal carbons, e.g., methandiyl (methano); ethan-1,2-diyl(ethano); propan-1,3-diyl (propano); butan-1,4-diyl (butano); and thelike (also referred to as alkylenos, defined infra).

“Alkdiyl” by itself or as part of another substituent refers to asaturated or unsaturated, branched, straight-chain or cyclic divalenthydrocarbon group having the stated number of carbon atoms (i.e., C1-C6means from one to six carbon atoms) derived by the removal of onehydrogen atom from each of two different carbon atoms of a parentalkane, alkene or alkyne, or by the removal of two hydrogen atoms from asingle carbon atom of a parent alkane, alkene or alkyne. The twomonovalent radical centers or each valency of the divalent radicalcenter can form bonds with the same or different atoms. Typical alkdiylgroups include, but are not limited to methandiyl; ethyldiyls such asethan-1,1-diyl, ethan-1,2-diyl, ethen-1,1-diyl, ethen-1,2-diyl;propyldiyls such as propan-1,1-diyl, propan-1,2-diyl, propan-2,2-diyl,propan-1,3-diyl, cyclopropan-1,1-diyl, cyclopropan-1,2-diyl,prop-1-en-1,1-diyl, prop-1-en-1,2-diyl, prop-2-en-1,2-diyl,prop-1-en-1,3-diyl, cycloprop-1-en-1,2-diyl, cycloprop-2-en-1,2-diyl,cycloprop-2-en-1,1-diyl-, prop-1-yn-1,3-diyl, etc.; butyldiyls such as,butan-1,1-diyl, butan-1,2-diyl, butan-1,3-diyl, butan-1,4-diyl,butan-2,2-diyl, 2-methyl-propan-1,1-diyl, 2-methyl-propan-1,2-diyl,cyclobutan-1,1-diyl; cyclobutan-1,2-diyl, cyclobutan-1,3-diyl,but-1-en-1,1-diyl, but-1-en-1,2-diyl, but-1-en-1,3-diyl,but-1-en-1,4-diyl, 2-methyl-prop-1-en-1,1-diyl,2-methanylidene-propan-1,1-diyl, buta-1,3-dien-1,1-diyl,buta-1,3-dien-1,2-diyl, buta-1,3-dien-1,3-diyl, buta-1,3-dien-1,4-diyl,cyclobut-1-en-1,2-diyl, cyclobut-1-en-1,3-diyl, cyclobut-2-en-1,2-diyl,cyclobuta-1,3-dien-1,2-diyl, cyclobuta-1,3-dien-1,3-diyl,but-1-yn-1,3-diyl, but-1-yn-1,4-diyl, buta-1,3-diyn-1,4-diyl, etc.; andthe like. Where specific levels of saturation are intended, thenomenclature alkandiyl, alkendiyl and/or alkyndiyl is used. In apreferred embodiment, the alkdiyl group is (C1-C6)alkdiyl. Alsopreferred are saturated acyclic alkanyldiyl groups in which the radicalcenters are at the terminal carbons, e.g., methandiyl (methano);ethan-1,2-diyl (ethano); propan-1,3-diyl (propano); butan-1,4-diyl(butano); and the like (also referred to as alkylenes, defined infra)

“Alkyleno” by itself or as part of another substituent refers to astraight-chain saturated or unsaturated alkyldiyl group having twoterminal monovalent radical centers derived by the removal of onehydrogen atom from each of the two terminal carbon atoms ofstraight-chain parent alkane, alkene or alkyne. The locant of a doublebond or triple bond, if present, in a particular alkyleno is indicatedin square brackets. Typical alkyleno groups include, but are not limitedto, methano; ethylenos such as ethano, etheno, ethyno; propylenos suchas propano, prop[1]eno, propa[1,2]dieno, prop[1]yno, etc.; butylenossuch as butano, but[1]eno, but[2]eno, buta[1,3]dieno, but[1]yno,but[2]yno, buta[1,3]diyno, etc.; and the like. Where specific levels ofsaturation are intended, the nomenclature alkano, alkeno and/or alkynois used. In preferred embodiments, the alkyleno group is (C1-C6) or(C1-C3)alkyleno. Also preferred are straight-chain saturated alkanogroups, e.g., methano, ethano, propano, butano, and the like.

“Heteroalkyl,” “Heteroalkanyl,” “Heteroalkenyl,” “Heteroalkynyl,”“Heteroalkyldiyl” and “Heteroalkyleno” by themselves or as part ofanother substituent refer to alkyl, alkanyl, alkenyl, alkynyl, alkyldiyland alkyleno groups, respectively, in which one or more of the carbonatoms are each independently replaced with the same or differentheteratoms or heteroatomic groups. Typical heteroatoms and/orheteroatomic groups which can replace the carbon atoms include, but arenot limited to, —O—, —S—, —S—O—, —NR′—, —PH—, —S(O)—, —S(O)₂—,—S(O)NR′—, —S(O)₂NR′—, and the like, including combinations thereof,where each R′ is independently hydrogen or (C1-C6)alkyl.

“Cycloalkyl” and “Heterocycloalkyl” by themselves or as part of anothersubstituent refer to cyclic versions of “alkyl” and “heteroalkyl”groups, respectively. For heteroalkyl groups, a heteroatom can occupythe position that is attached to the remainder of the molecule. Typicalcycloalkyl groups include, but are not limited to, cyclopropyl;cyclobutyls such as cyclobutanyl and cyclobutenyl; cyclopentyls such ascyclopentanyl and cyclopentenyl; cyclohexyls such as cyclohexanyl andcyclohexenyl; and the like. Typical heterocycloalkyl groups include, butare not limited to, tetrahydrofuranyl (e.g., tetrahydrofuran-2-yl,tetrahydrofuran-3-yl, etc.), piperidinyl (e.g., piperidin-1-yl,piperidin-2-yl, etc.), morpholinyl (e.g., morpholin-3-yl,morpholin-4-yl, etc.), piperazinyl (e.g., piperazin-1-yl,piperazin-2-yl, etc.), and the like.

“Acyclic Heteroatomic Bridge” refers to a divalent bridge in which thebackbone atoms are exclusively heteroatoms and/or heteroatomic groups.Typical acyclic heteroatomic bridges include, but are not limited to,—O—, —S—, —S—O—, —NR′—, —PH—, —S(O)—, —S(O)₂—, —S(O)NR′—, —S(O)₂NR′—,and the like, including combinations thereof, where each R′ isindependently hydrogen or (C1-C6)alkyl.

“Parent Aromatic Ring System” refers to an unsaturated cyclic orpolycyclic ring system having a conjugated π electron system.Specifically included within the definition of “parent aromatic ringsystem” are fused ring systems in which one or more of the rings arearomatic and one or more of the rings are saturated or unsaturated, suchas, for example, fluorene, indane, indene, phenalene,tetrahydronaphthalene, etc. Typical parent aromatic ring systemsinclude, but are not limited to, aceanthrylene, acenaphthylene,acephenanthrylene, anthracene, azulene, benzene, chrysene, coronene,fluoranthene, fluorene, hexacene, hexaphene, hexylene, indacene,s-indacene, indane, indene, naphthalene, octacene, octaphene, octalene,ovalene, penta-2,4-diene, pentacene, pentalene, pentaphene, perylene,phenalene, phenanthrene, picene, pleiadene, pyrene, pyranthrene,rubicene, tetrahydronaphthalene, triphenylene, trinaphthalene, and thelike, as well as the various hydro isomers thereof.

“Aryl” by itself or as part of another substituent refers to amonovalent aromatic hydrocarbon group having the stated number of carbonatoms (i.e., C5-C15 means from 5 to 15 carbon atoms) derived by theremoval of one hydrogen atom from a single carbon atom of a parentaromatic ring system. Typical aryl groups include, but are not limitedto, groups derived from aceanthrylene, acenaphthylene,acephenanthrylene, anthracene, azulene, benzene, chrysene, coronene,fluoranthene, fluorene, hexacene, hexaphene, hexylene, as-indacene,s-indacene, indane, indene, naphthalene, octacene, octaphene, octalene,ovalene, penta-2,4-diene, pentacene, pentalene, pentaphene, perylene,phenalene, phenanthrene, picene, pleiadene, pyrene, pyranthrene,rubicene, triphenylene, trinaphthalene, and the like, as well as thevarious hydro isomers thereof. In preferred embodiments, the aryl groupis (C5-C15)aryl, with (C5-C10) being even more preferred. Particularlypreferred aryls are cyclopentadienyl, phenyl and naphthyl.

“Arylaryl” by itself or as part of another substituent refers to amonovalent hydrocarbon group derived by the removal of one hydrogen atomfrom a single carbon atom of a ring system in which two or moreidentical or non-identical parent aromatic ring systems are joineddirectly together by a single bond, where the number of such direct ringjunctions is one less than the number of parent aromatic ring systemsinvolved. Typical arylaryl groups include, but are not limited to,biphenyl, triphenyl, phenyl-naphthyl, binaphthyl, biphenyl-naphthyl, andthe like. Where the number of carbon atoms in an arylaryl group arespecified, the numbers refer to the carbon atoms comprising each parentaromatic ring. For example, (C5-C15)arylaryl is an arylaryl group inwhich each aromatic ring comprises from 5 to 15 carbons, e.g., biphenyl,triphenyl, binaphthyl, phenylnaphthyl, etc. Preferably, each parentaromatic ring system of an arylaryl group is independently a (C5-C15)aromatic, more preferably a (C5-C10) aromatic. Also preferred arearylaryl groups in which all of the parent aromatic ring systems areidentical, e.g., biphenyl, triphenyl, binaphthyl, trinaphthyl, etc.

“Biaryl” by itself or as part of another substituent refers to anarylaryl group having two identical parent aromatic systems joineddirectly together by a single bond. Typical biaryl groups include, butare not limited to, biphenyl, binaphthyl, bianthracyl, and the like.Preferably, the aromatic ring systems are (C5-C15) aromatic rings, morepreferably (C5-C10) aromatic rings. A particularly preferred biarylgroup is biphenyl.

“Arylalkyl” by itself or as part of another substituent refers to anacyclic alkyl group in which one of the hydrogen atoms bonded to acarbon atom, typically a terminal or sp³ carbon atom, is replaced withan aryl group. Typical arylalkyl groups include, but are not limited to,benzyl, 2-phenylethan-1-yl, 2-phenylethen-1-yl, naphthylmethyl,2-naphthylethan-1-yl, 2-naphthylethen-1-yl, naphthobenzyl,2-naphthophenylethan-1-yl and the like. Where specific alkyl moietiesare intended, the nomenclature arylalkanyl, arylakenyl and/orarylalkynyl is used. In preferred embodiments, the arylalkyl group is(C6-C21)arylalkyl, e.g., the alkanyl, alkenyl or alkynyl moiety of thearylalkyl group is (C1-C6) and the aryl moiety is (C5-C15). Inparticularly preferred embodiments the arylalkyl group is (C6-C13),e.g., the alkanyl, alkenyl or alkynyl moiety of the arylalkyl group is(C1-C3) and the aryl moiety is (C5-C10).

“Parent Heteroaromatic Ring System” refers to a parent aromatic ringsystem in which one or more carbon atoms are each independently replacedwith the same or different heteroatoms or heteroatomic groups. Typicalheteroatoms or heteroatomic groups to replace the carbon atoms include,but are not limited to, N, NH, P, O, S, S(O), S(O)₂, Si, etc.Specifically included within the definition of “parent heteroaromaticring systems” are fused ring systems in which one or more of the ringsare aromatic and one or more of the rings are saturated or unsaturated,such as, for example, benzodioxan, benzofuran, chromane, chromene,indole, indoline, xanthene, etc. Also included in the definition of“parent heteroaromatic ring system” are those recognized rings thatinclude common substituents, such as, for example, benzopyrone and1-methyl-1,2,3,4-tetrazole. Typical parent heteroaromatic ring systemsinclude, but are not limited to, acridine, benzimidazole, benzisoxazole,benzodioxan, benzodioxole, benzofuran, benzopyrone, benzothiadiazole,benzothiazole, benzotriazole, benzoxaxine, benzoxazole, benzoxazoline,carbazole, β-carboline, chromane, chromene, cinnoline, furan, imidazole,indazole, indole, indoline, indolizine, isobenzofuran, isochromene,isoindole, isoindoline, isoquinoline, isothiazole, isoxazole,naphthyridine, oxadiazole, oxazole, perimidine, phenanthridine,phenanthroline, phenazine, phthalazine, pteridine, purine, pyran,pyrazine, pyrazole, pyridazine, pyridine, pyrimidine, pyrrole,pyrrolizine, quinazoline, quinoline, quinolizine, quinoxaline,tetrazole, thiadiazole, thiazole, thiophene, triazole, xanthene, and thelike.

“Heteroaryl” by itself or as part of another substituent refers to amonovalent heteroaromatic group having the stated number of ring atoms(e.g., “5-14 membered” means from 5 to 14 ring atoms) derived by theremoval of one hydrogen atom from a single atom of a parentheteroaromatic ring system. Typical heteroaryl groups include, but arenot limited to, groups derived from acridine, benzimidazole,benzisoxazole, benzodioxan, benzodiaxole, benzofuran, benzopyrone,benzothiadiazole, benzothiazole, benzotriazole, benzoxazine,benzoxazole, benzoxazoline, carbazole, β-carboline, chromane, chromene,cinnoline, furan, imidazole, indazole, indole, indoline, indolizine,isobenzofuran, isochromene, isoindole, isoindoline, isoquinoline,isothiazole, isoxazole, naphthyridine, oxadiazole, oxazole, perimidine,phenanthridine, phenanthroline, phenazine, phthalazine, pteridine,purine, pyran, pyrazine, pyrazole, pyridazine, pyridine, pyrimidine,pyrrole, pyrrolizine, quinazoline, quinoline, quinolizine, quinoxaline,tetrazole, thiadiazole, thiazole, thiophene, triazole, xanthene, and thelike, as well as the various hydro isomers thereof. In preferredembodiments, the heteroaryl group is a 5-14 membered heteroaryl, with5-10 membered heteroaryl being particularly preferred.

“Heteroaryl-Heteroaryl” by itself or as part of another substituentrefers to a monovalent heteroaromatic group derived by the removal ofone hydrogen atom from a single atom of a ring system in which two ormore identical or non-identical parent heteroaromatic ring systems arejoined directly together by a single bond, where the number of suchdirect ring junctions is one less than the number of parentheteroaromatic ring systems involved. Typical heteroaryl-heteroarylgroups include, but are not limited to, bipyridyl, tripyridyl,pyridylpurinyl, bipurinyl, etc. Where the number of atoms are specified,the numbers refer to the number of atoms comprising each parentheteroaromatic ring systems. For example, 5-15 memberedheteroaryl-heteroaryl is a heteroaryl-heteroaryl group in which eachparent heteroaromatic ring system comprises from 5 to 15 atoms, e.g.,bipyridyl, tripuridyl, etc. Preferably, each parent heteroaromatic ringsystem is independently a 5-15 membered heteroaromatic, more preferablya 5-10 membered heteroaromatic. Also preferred are heteroaryl-heteroarylgroups in which all of the parent heteroaromatic ring systems areidentical.

“Biheteroaryl” by itself or as part of another substituent refers to aheteroaryl-heteroaryl group having two identical parent heteroaromaticring systems joined directly together by a single bond. Typicalbiheteroaryl groups include, but are not limited to, bipyridyl,bipurinyl, biquinolinyl, and the like. Preferably, the heteroaromaticring systems are 5-15 membered heteroaromatic rings, more preferably5-10 membered heteroaromatic rings.

“Heteroarylalkyl” by itself or as part of another substituent refers toan acyclic alkyl group in which one of the hydrogen atoms bonded to acarbon atom, typically a terminal or sp³ carbon atom, is replaced with aheteroaryl group. Where specific alkyl moieties are intended, thenomenclature heteroarylalkanyl, heteroarylakenyl and/orheteroarylalkynyl is used. In preferred embodiments, the heteroarylalkylgroup is a 6-21 membered heteroarylalkyl, e.g., the alkanyl, alkenyl oralkynyl moiety of the heteroarylalkyl is (C1-C6)alkyl and the heteroarylmoiety is a 5-15-membered heteroaryl. In particularly preferredembodiments, the heteroarylalkyl is a 6-13 membered heteroarylalkyl,e.g., the alkanyl, alkenyl or alkynyl moiety is (C1-C3)alkyl and theheteroaryl moiety is a 5-10 membered heteroaryl.

“Halogen” or “Halo” by themselves or as part of another substituent,unless otherwise stated, refer to fluoro, chloro, bromo and iodo.

“Haloalkyl” by itself or as part of another substituent refers to analkyl group in which one or more of the hydrogen atoms is replaced witha halogen. Thus, the term “haloalkyl” is meant to includemonohaloalkyls, dihaloalkyls, trihaloalkyls, etc. up to perhaloalkyls.For example, the expression “(C1-C2)haloalkyl” includes fluoromethyl,difluoromethyl, trifluoromethyl, 1-fluoroethyl, 1,1-difluoroethyl,1,2-difluoroethyl, 1,1,1-trifluoroethyl, perfluoroethyl, etc.

The above-defined groups may include prefixes and/or suffixes that arecommonly used in the art to create additional well-recognizedsubstituent groups. As examples, “alkyloxy” or “alkoxy” refers to agroup of the formula —OR″, “alkylamine” refers to a group of the formula—NHR″ and “dialkylamine” refers to a group of the formula —NR″R″, whereeach R″ is independently an alkyl. As another example, “haloalkoxy” or“haloalkyloxy” refers to a group of the formula —OR′″, where R′″ is ahaloalkyl.

The present invention is also drawn to methods for treating arterialinflammation, arthritis, psoriasis, urticara, vasculitis, asthma, ocularinflammation, pulmonary inflammation, pulmonary fibrosis, seborrheicdermatitis, pustular dermatosis, or cardiovascular diseases in a subjectby administration of one or more of the DHA analogs described herein.Disease states or conditions that are associated with inflammation suchas the recruitment of neutrophils, leukocytes and/or cytokines areincluded within the general scope of inflammation and include, forexample, Addiction, AIDS, Alcohol-related disorders, Allergy,Alzheimer's disease, Anesthesiology, Anti-infectives, Anti-inflammatoryagents, Arthritis, Asthma, Atherosclerosis, Bone diseases, Breastcancer, Cancer, Cardiovascular diseases, Child health, Colon cancer,Congenital defects, Decision analysis, Degenerative neurologicdisorders, Dementia, Dermatology, Diabetes mellitus, Diagnostics, Drugdelivery, Drug discovery/screen, Endocrine disorders, ENT, Epidemiology,Eye diseases, Fetal and maternal medicine, Gastrointestinal disorders,Gene therapy, Genetic diagnostics, Genetics, Genitourinary disorders,Geriatric medicine, Growth and Development, Hearing, Hematologicdisorders, Hepatobiliary disorders, Hypertension, Imaging, Immunology,Infectious diseases, Leukemia/lymphoma, Lung cancer, Metabolicdisorders, Neonatology, Neurological disorders, Neuromuscular disorders,Nuclear medicine, Obesity/eating disorders, Orthopedic, Other, Parasiticdiseases, Perinatal disorders, Pregnancy, Preventative medicine,Prostate cancer, Psychiatric disorders, Pulmonary disorders, Radiology,Renal disorders, Reproduction, Rheumatic diseases, Stroke, Surgical,Transplantation, Vaccines, Vascular medicine, Wound healing, oralinfections, periodontal disease, brain injury, trauma and neuronalinflammation, and Women's health.

The pharmaceutical compositions of the invention include a“therapeutically effective amount” or a “prophylactically effectiveamount” of one or more of the DHA analogs of the invention. A“therapeutically effective amount” refers to an amount effective, atdosages and for periods of time necessary, to achieve the desiredtherapeutic result, e.g., a diminishment or prevention of effectsassociated with various disease states or conditions. A therapeuticallyeffective amount of the DHA analog may vary according to factors such asthe disease state, age, sex, and weight of the individual, and theability of the therapeutic compound to elicit a desired response in theindividual. A therapeutically effective amount is also one in which anytoxic or detrimental effects of the therapeutic agent are outweighed bythe therapeutically beneficial effects.

A “prophylactically effective amount” refers to an amount effective, atdosages and for periods of time necessary, to achieve the desiredprophylactic result. Typically, since a prophylactic dose is used insubjects prior to or at an earlier stage of disease, theprophylactically effective amount will be less than the therapeuticallyeffective amount.

Dosage regimens may be adjusted to provide the optimum desired response(e.g., a therapeutic or prophylactic response). For example, a singlebolus may be administered, several divided doses may be administeredover time or the dose may be proportionally reduced or increased asindicated by the exigencies of the therapeutic situation. It isespecially advantageous to formulate parenteral compositions in dosageunit form for ease of administration and uniformity of dosage. Dosageunit form as used herein refers to physically discrete units suited asunitary dosages for the mammalian subjects to be treated; each unitcontaining a predetermined quantity of active compound calculated toproduce the desired therapeutic effect in association with the requiredpharmaceutical carrier. The specification for the dosage unit forms ofthe invention are dictated by and directly dependent on (a) the uniquecharacteristics of the DHA analog and the particular therapeutic orprophylactic effect to be achieved, and (b) the limitations inherent inthe art of compounding such an active compound for the treatment ofsensitivity in individuals.

An exemplary, non-limiting range for a therapeutically orprophylactically effective amount of a DHA analog of the invention is0.1-20 mg/kg, more preferably 1-10 mg/kg. It is to be noted that dosagevalues may vary with the type and severity of the condition to bealleviated. It is to be further understood that for any particularsubject, specific dosage regimens should be adjusted over time accordingto the individual need and the professional judgment of the personadministering or supervising the administration of the compositions, andthat dosage ranges set forth herein are exemplary only and are notintended to limit the scope or practice of the claimed composition.

When the compounds of the present invention are administered aspharmaceuticals, to humans and mammals, they can be given per se or as apharmaceutical composition containing, for example, 0.1 to 99.5% (morepreferably, 0.5 to 90%) of active ingredient, i.e., at least one DHAanalog, in combination with a pharmaceutically acceptable carrier.

In certain embodiments, the compounds of the present invention maycontain one or more acidic functional groups and, thus, are capable offorming pharmaceutically acceptable salts with pharmaceuticallyacceptable bases. The term “pharmaceutically acceptable salts, esters,amides, and prodrugs” as used herein refers to those carboxylate salts,amino acid addition salts, esters, amides, and prodrugs of the compoundsof the present invention which are, within the scope of sound medicaljudgment, suitable for use in contact with the tissues of patientswithout undue toxicity, irritation, allergic response, and the like,commensurate with a reasonable benefit/risk ratio, and effective fortheir intended use of the compounds of the invention. The term “salts”refers to the relatively non toxic, inorganic and organic acid additionsalts of compounds of the present invention. These salts can be preparedin situ during the final isolation and purification of the compounds orby separately reacting the purified compound in its free base form witha suitable organic or inorganic acid and isolating the salt thus formed.These may include cations based on the alkali and alkaline earth metals,such as sodium, lithium, potassium, calcium, magnesium and the like, aswell as non toxic ammonium, quaternary ammonium, and amine cationsincluding, but not limited to ammonium, tetramethylammonium,tetraethylammonium, methylamine, dimethylamine, trimethylamine,triethylamine, ethylamine, and the like. (See, for example, Berge S. M.,et al., “Pharmaceutical Salts,” J. Pharm. Sci., 1977; 66:1 19 which isincorporated herein by reference).

The term “pharmaceutically acceptable esters” refers to the relativelynon-toxic, esterified products of the compounds of the presentinvention. These esters can be prepared in situ during the finalisolation and purification of the compounds, or by separately reactingthe purified compound in its free acid form or hydroxyl with a suitableesterifying agent. Carboxylic acids can be converted into esters viatreatment with an alcohol in the presence of a catalyst. The term isfurther intended to include lower hydrocarbon groups capable of beingsolvated under physiological conditions, e.g., alkyl esters, methyl,ethyl and propyl esters.

Wetting agents, emulsifiers and lubricants, such as sodium laurylsulfate and magnesium stearate, as well as coloring agents, releaseagents, coating agents, sweetening, flavoring and perfuming agents,preservatives and antioxidants can also be present in the compositions.

Examples of pharmaceutically acceptable antioxidants include: watersoluble antioxidants, such as ascorbic acid, cysteine hydrochloride,sodium bisulfate, sodium metabisulfite, sodium sulfite and the like;oil-soluble antioxidants, such as ascorbyl palmitate, butylatedhydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propylgallate, alpha-tocopherol, and the like; and metal chelating agents,such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol,tartaric acid, phosphoric acid, and the like.

Formulations of the present invention include those suitable forintravenous, oral, nasal, topical, transdermal, buccal, sublingual,rectal, vaginal and/or parenteral administration. The formulations mayconveniently be presented in unit dosage form and may be prepared by anymethods well known in the art of pharmacy. The amount of activeingredient which can be combined with a carrier material to produce asingle dosage form will generally be that amount of the compound whichproduces a therapeutic effect. Generally, out of one hundred percent,this amount will range from about 1 percent to about ninety-nine percentof active ingredient, preferably from about 5 percent to about 70percent, most preferably from about 10 percent to about 30 percent.

Methods of preparing these formulations or compositions include the stepof bringing into association a compound of the present invention withthe carrier and, optionally, one or more accessory ingredients. Ingeneral, the formulations are prepared by uniformly and intimatelybringing into association a compound of the present invention withliquid carriers, or finely divided solid carriers, or both, and then, ifnecessary, shaping the product.

Formulations of the invention suitable for oral administration may be inthe finial of capsules, cachets, pills, tablets, lozenges (using aflavored basis, usually sucrose and acacia or tragacanth), powders,granules, or as a solution or a suspension in an aqueous or non-aqueousliquid, or as an oil-in-water or water-in-oil liquid emulsion, or as anelixir or syrup, or as pastilles (using an inert base, such as gelatinand glycerin, or sucrose and acacia) and/or as mouth washes and thelike, each containing a predetermined amount of a compound of thepresent invention as an active ingredient. A compound of the presentinvention may also be administered as a bolus, electuary or paste.

In solid dosage forms of the invention for oral administration(capsules, tablets, pills, dragees, powders, granules and the like), theactive ingredient is mixed with one or more pharmaceutically acceptablecarriers, such as sodium citrate or dicalcium phosphate, and/or any ofthe following: fillers or extenders, such as starches, lactose, sucrose,glucose, mannitol, and/or silicic acid; binders, such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone,sucrose and/or acacia; humectants, such as glycerol; disintegratingagents, such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate; solutionretarding agents, such as paraffin; absorption accelerators, such asquaternary ammonium compounds; wetting agents, such as, for example,cetyl alcohol and glycerol monostearate; absorbents, such as kaolin andbentonite clay; lubricants, such a talc, calcium stearate, magnesiumstearate, solid polyethylene glycols, sodium lauryl sulfate, andmixtures thereof; and coloring agents. In the case of capsules, tabletsand pills, the pharmaceutical compositions may also comprise bufferingagents. Solid compositions of a similar type may also be employed asfillers in soft and hard-filled gelatin capsules using such excipientsas lactose or milk sugars, as well as high molecular weight polyethyleneglycols and the like.

A tablet may be made by compression or molding, optionally with one ormore accessory ingredients. Compressed tablets may be prepared usingbinder (for example, gelatin or hydroxypropylmethyl cellulose),lubricant, inert diluent, preservative, disintegrant (for example,sodium starch glycolate or cross-linked sodium carboxymethyl cellulose),surface-active or dispersing agent. Molded tablets may be made bymolding in a suitable machine a mixture of the powdered compoundmoistened with an inert liquid diluent.

The tablets, and other solid dosage forms of the pharmaceuticalcompositions of the present invention, such as dragees, capsules, pillsand granules, may optionally be scored or prepared with coatings andshells, such as enteric coatings and other coatings well known in thepharmaceutical-formulating art. They may also be formulated so as toprovide slow or controlled release of the active ingredient thereinusing, for example, hydroxypropylmethyl cellulose in varying proportionsto provide the desired release profile, other polymer matrices,liposomes and/or microspheres. They may be sterilized by, for example,filtration through a bacteria-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved in sterile water, or some other sterile injectable mediumimmediately before use. These compositions may also optionally containopacifying agents and may be of a composition that they release theactive ingredient(s) only, or preferentially, in a certain portion ofthe gastrointestinal tract, optionally, in a delayed manner. Examples ofembedding compositions which can be used include polymeric substancesand waxes. The active ingredient can also be in micro-encapsulated form,if appropriate, with one or more of the above-described excipients.

Liquid dosage forms for oral administration of the compounds of theinvention include pharmaceutically acceptable emulsions, microemulsions,solutions, suspensions, syrups and elixirs. In addition to the activeingredient, the liquid dosage forms may contain inert diluents commonlyused in the art, such as, for example, water or other solvents,solubilizing agents and emulsifiers, such as ethyl alcohol, isopropylalcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzylbenzoate, propylene glycol, 1,3-butylene glycol, oils (in particular,cottonseed, groundnut, corn, germ, olive, castor and sesame oils),glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acidesters of sorbitan, and mixtures thereof.

Besides inert diluents, the oral compositions can also include adjuvantssuch as wetting agents, emulsifying and suspending agents, sweetening,flavoring, coloring, perfuming and preservative agents.

Suspensions, in addition to the active compounds, may contain suspendingagents as, for example, ethoxylated isostearyl alcohols, polyoxyethylenesorbitol and sorbitan esters, microcrystalline cellulose, aluminummetahydroxide, bentonite, agar-agar and tragacanth, and mixturesthereof.

Formulations of the pharmaceutical compositions of the invention forrectal or vaginal administration may be presented as a suppository,which may be prepared by mixing one or more compounds of the inventionwith one or more suitable nonirritating excipients or carrierscomprising, for example, cocoa butter, polyethylene glycol, asuppository wax or a salicylate, and which is solid at room temperature,but liquid at body temperature and, therefore, will melt in the rectumor vaginal cavity and release the active compound.

Formulations of the present invention which are suitable for vaginaladministration also include pessaries, tampons, creams, gels, pastes,foams or spray formulations containing such carriers as are known in theart to be appropriate.

Dosage forms for the topical or transdermal administration of a compoundof this invention include powders, sprays, ointments, pastes, creams,lotions, gels, solutions, patches and inhalants. The active compound maybe mixed under sterile conditions with a pharmaceutically acceptablecarrier, and with any preservatives, buffers, or propellants which maybe required.

The ointments, pastes, creams and gels may contain, in addition to anactive compound of this invention, excipients, such as animal andvegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulosederivatives, polyethylene glycols, silicones, bentonites, silicic acid,talc and zinc oxide, or mixtures thereof.

Powders and sprays can contain, in addition to a compound of thisinvention, excipients such as lactose, talc, silicic acid, aluminumhydroxide, calcium silicates and polyamide powder, or mixtures of thesesubstances. Sprays can additionally contain customary propellants, suchas chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons,such as butane and propane.

Transdermal patches have the added advantage of providing controlleddelivery of a compound of the present invention to the body. Such dosageforms can be made by dissolving or dispersing the compound in the propermedium. Absorption enhancers can also be used to increase the flux ofthe compound across the skin. The rate of such flux can be controlled byeither providing a rate controlling membrane or dispersing the activecompound in a polymer matrix or gel.

Ophthalmic formulations, eye ointments, powders, solutions and the like,are also contemplated as being within the scope of this invention. Suchsolutions are useful for the treatment of conjunctivitis.

Pharmaceutical compositions of this invention suitable for parenteraladministration comprise one or more compounds of the invention incombination with one or more pharmaceutically acceptable sterileisotonic aqueous or nonaqueous solutions, dispersions, suspensions oremulsions, or sterile powders which may be reconstituted into sterileinjectable solutions or dispersions just prior to use, which may containantioxidants, buffers, bacteriostats, solutes which render theformulation isotonic with the blood of the intended recipient orsuspending or thickening agents.

Examples of suitable aqueous and nonaqueous carriers which may beemployed in the pharmaceutical compositions of the invention includewater, ethanol, polyols (such as glycerol, propylene glycol,polyethylene glycol, and the like), and suitable mixtures thereof,vegetable oils, such as olive oil, and injectable organic esters, suchas ethyl oleate. Proper fluidity can be maintained, for example, by theuse of coating materials, such as lecithin, by the maintenance of therequired particle size in the case of dispersions, and by the use ofsurfactants.

These compositions may also contain adjuvants such as preservatives,wetting agents, emulsifying agents and dispersing agents. Prevention ofthe action of microorganisms may be ensured by the inclusion of variousantibacterial and antifungal agents, for example, paraben,chlorobutanol, phenol sorbic acid, and the like. It may also bedesirable to include isotonic agents, such as sugars, sodium chloride,and the like into the compositions. In addition, prolonged absorption ofthe injectable pharmaceutical form may be brought about by the inclusionof agents which delay absorption such as aluminum monostearate andgelatin.

In some cases, in order to prolong the effect of a drug, it is desirableto slow the absorption of the drug from subcutaneous or intramuscularinjection. This may be accomplished by the use of a liquid suspension ofcrystalline or amorphous material having poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolutionwhich, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally-administered drugform is accomplished by dissolving or suspending the drug in an oilvehicle.

Injectable depot forms are made by forming microencapsule matrices ofthe subject compounds in biodegradable polymers such aspolylactide-polyglycolide. Depending on the ratio of drug to polymer,and the nature of the particular polymer employed, the rate of drugrelease can be controlled. Examples of other biodegradable polymersinclude poly(orthoesters) and poly(anhydrides). Depot injectableformulations are also prepared by entrapping the drug in liposomes ormicroemulsions which are compatible with body tissue.

The preparations of the present invention may be given orally,parenterally, topically, or rectally. They are of course given by formssuitable for each administration route. For example, they areadministered in tablets or capsule form, by injection, inhalation, eyelotion, ointment, suppository, etc. administration by injection,infusion or inhalation; topical by lotion or ointment; and rectal bysuppositories. Intravenous injection administration is preferred.

The phrases “parenteral administration” and “administered parenterally”as used herein means modes of administration other than enteral andtopical administration, usually by injection, and includes, withoutlimitation, intravenous, intramuscular, intraarterial, intrathecal,intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal,transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular,subarachnoid, intraspinal and intrasternal injection and infusion.

The phrases “systemic administration,” “administered systematically,”“peripheral administration” and “administered peripherally” as usedherein mean the administration of a compound, drug or other materialother than directly into the central nervous system, such that it entersthe patient's system and, thus, is subject to metabolism and other likeprocesses, for example, subcutaneous administration.

These compounds may be administered to humans and other animals fortherapy by any suitable route of administration, including orally,nasally, as by, for example, a spray, rectally, intravaginally,parenterally, intracisternally and topically, as by powders, ointmentsor drops, including buccally and sublingually.

Regardless of the route of administration selected, the compounds of thepresent invention, which may be used in a suitable hydrated form, and/orthe pharmaceutical compositions of the present invention, are formulatedinto pharmaceutically acceptable dosage forms by conventional methodsknown to those of ordinary skill in the art. Actual dosage levels of theactive ingredients in the pharmaceutical compositions of this inventionmay be varied so as to obtain an amount of the active ingredient whichis effective to achieve the desired therapeutic response for aparticular patient, composition, and mode of administration, withoutbeing toxic to the patient.

The selected dosage level will depend upon a variety of factorsincluding the activity of the particular compound of the presentinvention employed, or the ester, salt or amide thereof, the route ofadministration, the time of administration, the rate of excretion of theparticular compound being employed, the duration of the treatment, otherdrugs, compounds and/or materials used in combination with theparticular compound employed, the age, sex, weight, condition, generalhealth and prior medical history of the patient being treated, and likefactors well known in the medical arts.

A physician or veterinarian having ordinary skill in the art can readilydetermine and prescribe the effective amount of the pharmaceuticalcomposition required. For example, the physician or veterinarian couldstart doses of the compounds of the invention employed in thepharmaceutical composition at levels lower than that required in orderto achieve the desired therapeutic effect and gradually increase thedosage until the desired effect is achieved.

In general, a suitable daily dose of a compound of the invention will bethat amount of the compound which is the lowest dose effective toproduce a therapeutic effect. Such an effective dose will generallydepend upon the factors described above. Generally, intravenous andsubcutaneous doses of the compounds of this invention for a patient,when used for the indicated analgesic effects, will range from about0.0001 to about 100 mg per kilogram of body weight per day, morepreferably from about 0.01 to about 50 mg per kg per day, and still morepreferably from about 0.1 to about 40 mg per kg per day. For example,between about 0.01 microgram and 20 micrograms, between about 20micrograms and 100 micrograms and between about 10 micrograms and 200micrograms of the compounds of the invention are administered per 20grams of subject weight.

If desired, the effective daily dose of the active compound may beadministered as two, three, four, five, six or more sub-dosesadministered separately at appropriate intervals throughout the day,optionally, in unit dosage forms.

The invention features an article of manufacture that contains packagingmaterial and DHA analog formulation contained within the packagingmaterial. This formulation contains an at least one DHA analog and thepackaging material contains a label or package insert indicating thatthe formulation can be administered to the subject to treat one or moreconditions as described herein, in an amount, at a frequency, and for aduration effective to treat or prevent such condition(s). Suchconditions are mentioned throughout the specification and areincorporated herein by reference. Suitable DHA analogs are describedherein.

The present invention surprisingly provides novel compounds,compositions and methods of use pertaining to dihydroxy analogues ofdocosahexaenoic acid (DHA) all having a hydroxyl group at C-14 of thecarbon chain and a second hydroxyl group at either the C-4, C-7 or C-14positions of the carbon chain. These materials are biogenically derivedand isolated from media.

In one embodiment, the invention pertains to a new and useful DHAanalogue such as a compound comprising the formula (I):

wherein each of P₁ and P₂ individually is a protecting group or ahydrogen atom;

wherein

is a double bond;

wherein Z is —C(O)OR^(d), —C(O)NR^(c)R^(c), —C(O)H, —C(NH)NR^(c)R^(c),—C(S)H, —C(S)OR^(d), —C(S)NR^(c)R^(c), or —CN;

each R^(a), is independently selected from hydrogen, (C1-C6)alkyl,(C3-C8)cycloalkyl, cyclohexyl, (C4-C11)cycloalkylalkyl, (C5-C10)aryl,phenyl, (C6-C16)arylalkyl, benzyl, 2-6 membered heteroalkyl, 3-8membered cycloheteroalkyl, morpholinyl, piperazinyl, homopiperazinyl,piperidinyl, 4-11 membered cycloheteroalkylalkyl, 5-10 memberedheteroaryl or 6-16 membered heteroarylalkyl;

each R^(c), is independently a protecting group or R^(a), or,alternatively, each R^(c) is taken together with the nitrogen atom towhich it is bonded to form a 5 to 8-membered cycloheteroalkyl orheteroaryl which may optionally include one or more of the same ordifferent additional heteroatoms and which may optionally be substitutedwith one or more of the same or different R^(a) or suitable R^(b)groups;

each R^(b) is independently selected from ═O, —OR^(d), (C1-C3), —OCF₃,═S, —SR^(d), ═NR^(d), ═NOR^(d), —NR^(c)R^(c), halogen, —CF₃, —CN, —NC,—OCN, —SCN, —NO, —NO₂, ═N₂, —N₃, —S(O)R^(d), —S(O)₂R^(d), —S(O)₂OR^(d),—S(O)NR^(c)R^(c), —S(O)₂NR^(c)R^(c), —OS(O)R^(d), —OS(O)₂R^(d),—OS(O)₂OR^(d), —OS(O)₂NR^(c)R^(c), —C(O)R^(d), —C(O)OR^(d),—C(O)NR^(c)R^(c), —C(NH)NR^(c)R^(c), —C(NH)NR^(c)R^(c), —C(NOH)R^(a),—C(NOH)NR^(c)R^(c), —OC(O)R^(d), —OC(O)OR^(d), —OC(O)NR^(c)R^(c),—OC(NH)NR^(c)R^(c), —OC(NR^(a))NR^(c)R^(c), —[NHC(O)]_(n)R^(d),—[NR^(a)C(O)]_(n)R^(d), —[NHC(O)]_(n)OR^(d), —[NR^(a)C(O)]_(n)R^(d),—[NHC(O)]_(n)NR^(c)R^(c), —[NR^(a)C(O)]_(n)NR^(c)R^(c),—[NHC(NH)]_(n)NR^(c)R^(c) or —[NR^(a)C(NR^(a))]_(n)NR^(c)R^(c);

each n, independently is an integer from 0 to 3; and

each R^(d), independently is a protecting group or R^(a);

or a pharmaceutically acceptable salt thereof, provided when Z is—C(O)OR, then R^(d) for Z is not a hydrogen. In certain aspects, P₁ andP₂ are both hydrogen atoms. In another aspect, the double bonds at the4, 10, 16 and 19 positions are each of Z configuration or the doublebonds at the 4, 16 and 19 positions are each of the Z configuration.

A particular isomer of interest of the DHA analogue (I) is (Ia)comprising the formula:

Another isomer of interest of the DHA analogue (I) is (Ib) comprisingthe formula:

referred to as the “double dioxygenation” product.

It should be understood that compounds (Ia) and (Ib) include allpharmaceutically acceptable salts, esters thereof, the purified/isolatedforms, as well as compounds wherein one or both of the hydroxyls areconverted into a protecting group as described herein.

In another aspect, the present invention provides new and useful DHAanalogues such as a purified compound comprising the formula (I):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d) and n are as previously defined. In oneaspect, P₁ and P₂ are both hydrogen atoms. In another aspect, Z is—C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In another embodiment,the double bonds at the 4, 10, 16 and 19 positions are each of Zconfiguration or the double bonds at the 4, 16 and 19 positions are eachof the Z configuration. In still another embodiment, the 7 hydroxyl hasan S configuration. In still yet another embodiment, the 14 hydroxyl hasan S configuration.

In another aspect, the present invention provides new and useful DHAanalogues such as a purified compound comprising the formula (Ic):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d) and n are as previously defined. R₁ isselected from (C1-C6)alkyl, (C3-C8)cycloalkyl, cyclohexyl,(C4-C11)cycloalkylalkyl, (C5-C10)aryl, phenyl, (C6-C16)arylalkyl,benzyl, 2-6 membered heteroalkyl, 3-8 membered cycloheteroalkyl,morpholinyl, piperazinyl, homopiperazinyl, piperidinyl, 4-11 memberedcycloheteroalkylalkyl, 5-10 membered heteroaryl or 6-16 memberedheteroarylalkyl. In one aspect, R₁ is a methyl group.

In one aspect, P₁ and P₂ are both hydrogen atoms. In another aspect, Zis —C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In another embodiment,the double bonds at the 4, 10, 16 and 19 positions are each of Zconfiguration or the double bonds at the 4, 16 and 19 positions are eachof the Z configuration. In still another embodiment, the 7 hydroxyl hasan S configuration. In still yet another embodiment, the 14 hydroxyl hasan S configuration.

In another aspect, the present invention provides new and useful DHAanalogues such as a purified compound comprising the formula (Id):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d) and n are as previously defined. R₂ isselected from (C1-C6)alkyl, (C3-C8)cycloalkyl, cyclohexyl,(C4-C11)cycloalkylalkyl, (C5-C10), phenyl, (C6-C16)arylalkyl, benzyl,2-6 membered heteroalkyl, 3-8 membered cycloheteroalkyl, morpholinyl,piperazinyl, homopiperazinyl, piperidinyl, 4-11 memberedcycloheteroalkylalkyl, 5-10 membered heteroaryl or 6-16 memberedheteroarylalkyl. In one aspect, R₂ is a methyl group.

In one aspect, P₁ and P₂ are both hydrogen atoms. In another aspect, Zis —C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In another embodiment,the double bonds at the 4, 10, 16 and 19 positions are each of Zconfiguration or the double bonds at the 4, 16 and 19 positions are eachof the Z configuration. In still another embodiment, the 7 hydroxyl hasan S configuration. In still yet another embodiment, the 14 hydroxyl hasan S configuration.

In another aspect, the present invention provides new and useful DHAanalogues such as a purified compound comprising the formula (Ie):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d), R₁, R₂ and n are as previously defined.

In one aspect, P₁ and P₂ are both hydrogen atoms. In another aspect, R₁and R₂ are both methyl groups. In still another aspect, Z is —C(O)OR^(d)and R^(d) of Z is a hydrogen atom. In another embodiment, the doublebonds at the 4, 10, 16 and 19 positions are each of Z configuration orthe double bonds at the 4, 16 and 19 positions are each of the Zconfiguration. In still another embodiment, the 7 hydroxyl has an Sconfiguration. In still yet another embodiment, the 14 hydroxyl has an Sconfiguration.

In another aspect, the invention pertains to a new and useful DHAanalogue such as a compound comprising the formula (II):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d) and n are as previously defined,provided when Z is —C(O)OR^(d), then R^(d) for Z is not a hydrogen. Inone embodiment, P₁ and P₂ are both hydrogen atoms. In anotherembodiment, the double bonds at the 7, 16 and 19 positions are each of Zconfiguration.

In another aspect, the present invention provides new and useful DHAanalogues such as a purified compound comprising the formula (II):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d) and n are as previously defined. In oneaspect, P₁ and P₂ are both hydrogen atoms. In another aspect, Z is—C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In still another aspect,the double bonds at the 7, 16 and 19 positions are each of Zconfiguration.

In another aspect, the present invention provides new and useful DHAanalogues such as a compound comprising the formula (IIa):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d), R₁ and n are as previously defined. Inone aspect, P₁ and P₂ are both hydrogen atoms. In another aspect, Z is—C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In still another aspect,the double bonds at the 7, 16 and 19 positions are each of Zconfiguration. In yet another aspect, R₁ is a methyl group.

In another aspect, the present invention provides new and useful DHAanalogues such as a compound comprising the formula (IIb):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d), R₂ and n are as previously defined. Inone aspect, P₁ and P₂ are both hydrogen atoms. In another aspect, Z is—C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In still another aspect,the double bonds at the 7, 16 and 19 positions are each of Zconfiguration. In yet another aspect, R₂ is a methyl group.

In another aspect, the present invention provides new and useful DHAanalogues such as a compound comprising the formula (IIc):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d), R₁, R₂ and n are as previously defined.In one aspect, P₁ and P₂ are both hydrogen atoms. In another aspect, Zis —C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In still anotheraspect, the double bonds at the 7, 16 and 19 positions are each of Zconfiguration. In yet another aspect, R₁ and R₂ are both methyl groups.

In another aspect, the present invention provides new and useful DHAanalogues such as a compound comprising formula (III):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d) and n are as previously defined,provided when Z is —C(O)OR^(d), then R^(d) for Z is not a hydrogen. Inone embodiment, P₁ and P₂ are both hydrogen atoms. In anotherembodiment, the double bonds at the 4, 7, 16 and 19 positions are eachof Z configuration.

In still another aspect, the present invention provides new and usefulDHA analogues such as a purified compound comprising the formula (III):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d) and n are as previously defined. In oneaspect, P₁ and P₂ are both hydrogen atoms. In another aspect, Z is—C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In another embodiment,the double bonds at the 4, 7, 16 and 19 positions are each of Zconfiguration.

In still another aspect, the present invention provides new and usefulDHA analogues such as a compound comprising the formula (IIIa):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d), R₁ and n are as previously defined. Inone aspect, P₁ and P₂ are both hydrogen atoms. In another aspect, Z is—C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In still another aspect,R₁ is a methyl group. In another embodiment, the double bonds at the 4,7, 16 and 19 positions are each of Z configuration.

In still another aspect, the present invention provides new and usefulDHA analogues such as a compound comprising the formula (IIIb):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d), R₂ and n are as previously defined. Inone aspect, P₁ and P₂ are both hydrogen atoms. In another aspect, Z is—C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In still another aspect,R₂ is a methyl group. In another embodiment, the double bonds at the 4,7, 16 and 19 positions are each of Z configuration.

In still another aspect, the present invention provides new and usefulDHA analogues such as a compound comprising the formula (IIIc):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d), R₁, R₂ and n are as previously defined.In one aspect, P₁ and P₂ are both hydrogen atoms. In another aspect, Zis —C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In still anotheraspect, R₁ and R₂ are both methyl groups. In another embodiment, thedouble bonds at the 4, 7, 16 and 19 positions are each of Zconfiguration.

In yet another aspect, the present invention provides new and useful DHAanalogues such as a compound comprising formula (IV):

wherein

, Z, R^(a), R^(b), R^(c), R^(d) and n are as previously defined,provided when Z is —C(O)OR^(d), then R^(d) for Z is not a hydrogen. Inone embodiment, the double bonds at the 4, 7, 16 and 19 positions areeach of Z configuration.

In still another aspect, the present invention provides new and usefulDHA analogues such as a purified compound comprising the formula (IV):

wherein

, Z, R^(a), R^(b), R^(c), R^(d) and n are as previously defined. In anaspect, Z is —C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In anotherembodiment, the double bonds at the 4, 7, 16 and 19 positions are eachof Z configuration.

In another aspect, the invention provides a compound comprising theformula (V):

wherein each of P₁, P₂ and P₃ individually is a protecting group or ahydrogen atom and

, Z, R^(a), R^(b), R^(c), R^(d) and n are as previously defined,provided when Z is —C(O)OR^(d), then R^(d) for Z is not a hydrogen. Inone aspect, P₁, P₂ and P₃ are all hydrogen atoms. In another aspect, thedouble bonds at the 7, 16 and 19 positions are each of Z configuration.

In still yet another aspect, the present invention provides a purifiedcompound comprising the formula (V):

wherein P₁, P₂, P₃,

, Z, R^(a), R^(b), R^(c), R^(d) and n are as previously defined. In oneaspect, P₁, P₂ and P₃ are all hydrogen atoms. In another aspect, Z is—C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In another embodiment,the double bonds at the 7, 16 and 19 positions are each of Zconfiguration.

In still yet another aspect, the present invention provides a compoundcomprising the formula (Va):

wherein P₁, P₂, P₃,

, Z, R^(a), R^(b), R^(c), R^(d), R₁ and n are as previously defined. Inone aspect, P₁, P₂ and P₃ are all hydrogen atoms. In another aspect, Zis —C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In another embodiment,the double bonds at the 7, 16 and 19 positions are each of Zconfiguration. In still another aspect, R₁ is a methyl group.

In another aspect, the present invention provides a compound comprisingthe formula (Vb):

wherein P₁, P₂, P₃,

, Z, R^(a), R^(b), R^(c), R^(d), R₂ and n are as previously defined. Inone aspect, P₁, P₂ and P₃ are all hydrogen atoms. In another aspect, Zis —C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In another embodiment,the double bonds at the 7, 16 and 19 positions are each of Zconfiguration. In still another aspect, R₂ is a methyl group.

In another aspect, the present invention provides a compound comprisingthe formula (Vc):

wherein P₁, P₂, P₃,

, Z, R^(a), R^(b), R^(c), R^(d), R₃ and n are as previously defined. Inone aspect, P₁, P₂ and P₃ are all hydrogen atoms. In another aspect, Zis —C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In another embodiment,the double bonds at the 7, 16 and 19 positions are each of Zconfiguration. In still another aspect, R₃ is a methyl group.

In another aspect, the present invention provides a compound comprisingthe formula (Vd):

wherein P₁, P₂, P₃,

, Z, R^(a), R^(b), R^(c), R^(d), R₁, R₂ and n are as previously defined.In one aspect, P₁, P₂ and P₃ are all hydrogen atoms. In another aspect,Z is —C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In anotherembodiment, the double bonds at the 7, 16 and 19 positions are each of Zconfiguration. In still another aspect, R₁ and R₂ are methyl groups.

In another aspect, the present invention provides a compound comprisingthe formula (Ve):

wherein P₁, P₂, P₃,

, Z, R^(a), R^(b), R^(c), R^(d), R₁, R₃ and n are as previously defined.In one aspect, P₁, P₂ and P₃ are all hydrogen atoms. In another aspect,Z is —C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In anotherembodiment, the double bonds at the 7, 16 and 19 positions are each of Zconfiguration. In still another aspect, R₁ and R₃ are methyl groups.

In another aspect, the present invention provides a compound comprisingthe formula (Vf):

wherein P₁, P₂, P₃,

, Z, R^(a), R^(b), R^(c), R^(d), R₂, R₃ and n are as previously defined.In one aspect, P₁, P₂ and P₃ are all hydrogen atoms. In another aspect,Z is —C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In anotherembodiment, the double bonds at the 7, 16 and 19 positions are each of Zconfiguration. In still another aspect, R₂ and R₃ are methyl groups.

In another aspect, the present invention provides a compound comprisingthe formula (Vg):

wherein P₁, P₂, P₃,

, Z, R^(a), R^(b), R^(c), R^(d), R_(I), R₂, R₃ and n are as previouslydefined. In one aspect, P₁, P₂ and P₃ are all hydrogen atoms. In anotheraspect, Z is —C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In anotherembodiment, the double bonds at the 7, 16 and 19 positions are each of Zconfiguration. In still another aspect, R₁, R₂ and R₃ are all methylgroups.

In another aspect, the present invention provides new and useful DHAanalogues such as a purified compound comprising the formula (VI):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d) and n are as previously defined.

In one aspect, P₁ and P₂ are both hydrogen atoms. In another aspect, Zis —C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In another embodiment,the double bonds at the 4, 10, 16 and 19 positions are each of Zconfiguration or the double bonds at the 4, 16 and 19 positions are eachof the Z configuration. In still another embodiment, the 7 hydroxyl hasan S configuration. In still yet another embodiment, the 14 hydroxyl hasan S configuration.

In another aspect, the present invention provides new and useful DHAanalogues such as a compound comprising the formula (VIa):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d), R₁ and n are as previously defined. Inone aspect, R₁ is a methyl group.

In one aspect, P₁ and P₂ are both hydrogen atoms. In another aspect, Zis —C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In another embodiment,the double bonds at the 4, 10, 16 and 19 positions are each of Zconfiguration or the double bonds at the 4, 16 and 19 positions are eachof the Z configuration. In still another embodiment, the 7 hydroxyl hasan S configuration. In still yet another embodiment, the 14 hydroxyl hasan S configuration.

In another aspect, the present invention provides new and useful DHAanalogues such as a compound comprising the formula (VIb):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d), R₂ and n are as previously defined. Inone aspect, R₂ is a methyl group.

In one aspect, P₁ and P₂ are both hydrogen atoms. In another aspect, Zis —C(O)OR^(d) and R^(d) of Z is a hydrogen atom. In another embodiment,the double bonds at the 4, 10, 16 and 19 positions are each of Zconfiguration or the double bonds at the 4, 16 and 19 positions are eachof the Z configuration. In still another embodiment, the 7 hydroxyl hasan S configuration. In still yet another embodiment, the 14 hydroxyl hasan S configuration.

In another aspect, the present invention provides new and useful DHAanalogues such as a compound comprising the formula (VIc):

wherein P₁, P₂,

, Z, R^(a), R^(b), R^(c), R^(d), R₁, R₂ and n are as previously defined.

In one aspect, P₁ and P₂ are both hydrogen atoms. In another aspect, R₁and R₂ are both methyl groups. In still another aspect, Z is —C(O)OR^(d)and R^(d) of Z is a hydrogen atom. In another embodiment, the doublebonds at the 4, 10, 16 and 19 positions are each of Z configuration orthe double bonds at the 4, 16 and 19 positions are each of the Zconfiguration. In still another embodiment, the 7 hydroxyl has an Sconfiguration. In still yet another embodiment, the 14 hydroxyl has an Sconfiguration.

In another aspect, the C-14 alcohol has an S configuration for thecompounds noted throughout the application.

It should be understood that the intermediates described herein are alsoincluded as part of the invention and can be considered active agents aswell. For example, ketone containing intermediates are within the scopeof the active agents as well as alkyne intermediates as describedherein.

Results and Discussion

Targeted Lipidomics of Resolving Inflammatory Exudates.

In view of the actions of specialized chemical mediators in resolution(6, 7), 17-HDHA was monitored as a biomarker of activation andconversion of endogenous DHA, as well as employed targeted lipidomics toquery whether other pathways were operative (see FIG. 1). During thiscourse of peritonitis, PMN rapidly entered, reaching max within 12 h. Inthis self-resolving system (19), PMN declined and were lost fromexudates, thus defining resolution (FIG. 1). Unbiased targeted mediatorlipidomics employing LC/MS/MS-based analyses were carried out with theseexudates. In addition to 17S-HDHA, a marker of resolvin and protectinbiosynthesis (6), endogenous DHA was converted to14S-hydroxydocosahexaenoic acid (14S-HDHA). Neither product wasidentified in exudates obtained with the lipoxygenase (LOX) inhibitoresculetin (n=3), and both were substantially reduced in peritonitislavages from 12/15-LOX-deficient mice (n=2, d=4). The appearance of14S-HDHA in this system accompanied 17-HDHA throughout the 72 h course,indicating that 14S-HDHA accumulated within the resolution phase.

These two LOX products were identified by characteristic diagnostic ionsin their respective mass spectra. FIG. 1B shows a representativespectrum of 17-HDHA formed in the time course, and 1C the spectrum of14S-HDHA as well as diagnostic ions for identification. These includedm/z 205, 138, and 233 specific for 14S-HDHA. The m/z 343 [M-H], m/z 325,299, and 281 ions are shared between 14-hydroxy- and17-hydroxydocosahexaenoic acid (see insets in FIGS. 1 B and C). Both14S-HDHA and 17-HDHA were also generated from endogenous DHA withisolated murine MΦs activated with Ca²⁺ ionophore A₂₃₁₈₇ (5.0 μM, pH7.45, n=4; representative values: 14S-HDHA, 325 pg/10⁶ cells; 17-HDHA,439 pg/10⁶ cells). These results demonstrate that during the time courseof acute inflammation and its resolution a sustained production ofendogenous LOX products was present in the initial acute phase at 2-4 hthat accumulated during resolution with highest levels of14S-HDHA>17-HDHA at 72 h.

It was determined that 14S-HDHA might be a marker reflecting activationof a novel DHA (C22:6) carbon 14-lipoxygenation pathway. This could leadto production of bioactive mediators via DHA, since monohydroxy productsof polyunsaturated fatty acids are biomarkers of pathways leading topotent bioactive molecules, as in the case of 17-HDHA and 17-HpDHA,precursors to resolvins and protectins (6, 20, 21). Also, 5-HETE is awell appreciated marker of arachidonic acid conversion to leukotrienes(2). To this end, resident peritoneal MΦs were isolated (see FIG. 2A,FACS inset), containing ˜85-90% MΦs and 10-15% lymphocytes, and14S-HpDHA was prepared via biogenic synthesis and incubated with thesecells to determine whether it was a precursor to new bioactive products.14S-HpDHA was isolated from 12-LOX incubated with HPLC-purified DHA(n=7) and was >98% S configuration determined by chiral LC/MS/MS. Both145-HpDHA (10 μM) and DHA (10 μM) were converted by resident MΦs to newproducts identified via LC/MS/MS-based mediator lipidomics (FIG. 2A)labeled I and II. Each had conjugated triene-containing UV chromophores,chromatographic behavior, and mass spectra consistent with7,14-dihydroxy-containing products with a C22 backbone originating fromDHA. Both gave essentially the same mass spectrum yet differentretention times, indicating that they were very likely isomers (FIGS. 2Band 2C). These were isolated and subject to gas chromatography-massspectrometry in order to further identify and confirm fragmentassignments and positional sites of oxygenation, e.g., carbon positionsof alcohol groups. GC-MS analyses confirmed assigned ions from LC/MS/MSand were consistent with 7,14-dihydroxy-containing productsbiosynthesized from DHA (FIG. 10). Isolated human MΦs incubated with14S-HpDHA also gave the novel 7,14-dihydroxy-containing product andmatched the compound from murine MΦs (n=3).

Novel Anti-Inflammatory and Pro-Resolving Mediators.

In parallel with these determinations, materials obtained from murineMΦs were assessed for potential bioactivity following extraction and C18solid-phase chromatography. MΦ-derived material showed remarkableanti-inflammatory properties (FIG. 3A) regulating PMN entry intozymosan-induced peritonitis when directly compared to other ω-3 fattyacid-derived mediators neuroprotectin/protectin D1 (20) and RvE1 (5, 6,14). Thus, these findings suggested that within MΦ isolates, obtainedfrom cells incubated with14S-hydroperoxydocosa-4Z,7Z,10Z,12E,16Z,19Z-hexaenoic acid, potentbioactive materials were biosynthesized from this precursor thatregulated PMN, preventing their infiltration. These substances werelikely to be very potent because only nanogram quantities given permouse elicited anti-inflammatory actions.

Given the ability of DHA to serve as a substrate for LOX forming bothdouble dioxygenation products related to protectin D1 as well as thecarbon 17-hydroperoxide-containing precursor of epoxide (20), it wasdetermined that 14S-HpDHA might also be substrate for doubledioxygenation. Sequential actions of 12-LOX and 5-LOX with DHA generated7S,14S-dihydroxydocosa-4Z,8E,10Z,12E,16Z,19Z-hexaenoic acid (n=5; FIG.2, dotted profile and FIG. 10). Likely this is an isomer of theMΦ-isolated material (FIGS. 2 and 3), because this reference compound(dotted curve in FIG. 2A) did not co-elute with MΦ-derived productsbeneath labels I and II. This double dioxygenation product at 0.1 ngdose per mouse reduced PMN infiltration in zymosan-induced peritonitisbut appeared to be less potent than the MΦ isolated material (FIG. 3B).Thus, results in FIG. 3 clearly demonstrate potent bioactions of thenovel 7,14-dihydroxy-containing product HPLC purified from resident MΦincubations with zymosan and14S-hydroperoxydocosa-4Z,7Z,10Z,12E,16Z,19Z-hexaenoic acid.

Further experiments were carried out to isolate material beneath peaks Iand II (FIG. 2A) and determine whether compound I is an isomer of II.Subjecting the isolated compounds to isomerization conditions (22)demonstrated that compound II likely contained a cis doublebond-containing triene structure sensitive to conversion to anall-trans-containing conjugated triene (i.e., 8E,10E,12E) isomer I inbench and work-up conditions. In view of ¹⁸O incorporation and epoxidetrapping (vide infra), it is likely that the isomer I peak alsocontained R/S racemates at carbon 7. Further RP-HPLC purification of the7,14-dihydroxy-containing product from MΦs was carried out to assess itsactions. Results in FIG. 3C demonstrate that at doses as low as 0.2ng/mouse the new compound potently reduced infiltration of PMN intosites of inflammation.

Macrophage-Enhanced Phagocytosis.

A key feature of a pro-resolving mediator, in addition to limiting PMNentry, is the dual action of stimulating MΦ uptake of apoptotic PMNand/or zymosan to stimulate resolution and microbial clearance (4, 13,23). Next it was determined whether the new MΦ-derived compound enhancedphagocytosis and compared it to RvE1, PD1 and an eicosanoid, PGE₂ (FIG.3D, inset). RvE1 and PD1 are potent enhancers of MΦ phagocytosis atconcentrations as low as 1 nM (13). For direct comparison, the doubledioxygenation product (7S,14S-diHDHA) was active but less potent thanthe new product isolated from MΦs. Thus, given its potent actions andnovel structure, the MΦ product was denoted maresin 1 (MaR1).

Maresin Biosynthetic Pathway.

FIG. 5 illustrates a hypothetical scheme for the maresin pathway. DHA isconverted to 14-hydroperoxydocosahexaenoic acid, likely via 12-LOX inhumans as shown in incubations of DHA and 12-LOX, followed by eitherreduction to 14S-HDHA and/or, via double dioxygenation, for examplesequential 12-LOX-5-LOX, to generate 7S,14S-diHDHA. In12/15-LOX-deficient mice, 14S-HDHA generation in peritonitis wasreduced >95%. The key 14S-hydroperoxide intermediate is enzymaticallyconverted to a 13(14)-epoxide-containing intermediate that is thenenzymatically hydrolyzed via a carbonium cation to bioactive7,14S-dihydroxydocosahexaenoic acid by creating a conjugated trienewithin three of the six double bonds. The results from oxygen-18 isotopeincorporation using H₂ ¹⁸O demonstrated >75% of the oxygen at carbon 7position was derived from H₂O (see FIG. 6) and not from molecularoxygen, as would have been the case if this 7 position alcohol groupwere generated by double lipoxygenation mechanism (cf. 20). In parallel,alcohol trapping with excess acidic methanol was carried out withisolated MΦs that gave methoxy-trapping product7-methoxy-14-hydroxydocosa-4Z,8,10,12,16Z,19Z-hexaenoic acid identifiedusing LC/MS/MS targeted profiling. Its MS² spectrum showed ionsconsistent with acid-assisted attack at carbon-7 position and additionof the methoxy, giving the MS² spectrum of m/z 373 at 10.2 min (FIG. 4inset for ion assignments, and FIG. 5). It is also possible that amethoxy addition could have occurred at carbon 13 position that couldgive essentially the same ions in MS². It is more likely that methoxyaddition was at carbon 7, because it is the least sterically hinderedend of the conjugated carbonium cation.

To provide further confirmation of the maresin pathway and biosyntheticscheme, isolated murine MΦs (15.5×10⁶ cells/3 ml, 37° C., 30 min) wereincubated with zymosan and deuterium-labeled DHA-d₅ (10 μM) containingfive deuterium atoms at 21, 21, 22, 22, and 22-carbon positions. MΦsconverted DHA-d₅ to 14S-HDHA-d₅, with diagnostic ions in its massspectrum at m/z 348, 330, 304, 286, and 205, as well as further evidencefor the 7,14S-dihydroxy-containing product. The dihydroxy structurecarried d₅, from precursor, and was confirmed with ions at m/z 364, 346,320, and 302, and the hydroxy groups' 7- and 14-positions supported bym/z 251, 223 and 250, 221 respectively (not shown). Together theseresults provide support for the biosynthesis of a 13(14) epoxideintermediate by MΦs from14S-hydroperoxydocosa-4Z,7Z,10Z,12E,16Z,19Z-hexaenoic acid (14S-HpDHA)that is enzymatically converted to the potent bioactive7,14-dihydroxydocosa-4Z,8,10,12,16Z,19Z-hexaenoic acid MaR1 in FIG. 5.

DHA (C22:6) is an essential fatty acid and member of the n-3 family offatty acids, which are in high levels in marine oils. It is essentialfor mammalian systems in that it is not biosynthesized de novo andtherefore a nutritional requirement (10, 11, 21). A novel pathway hereinis provided such that during resolution DHA is converted to new, potentbioactive products. This new pathway was identified using targetedmediator lipidomics of exudates from murine peritonitis and wasdemonstrated with both murine and human MΦs. Parallel biofunction andspectral analyses confirmed the new structures as7,14-dihydroxy-containing products biosynthesized from DHA. One of thenovel products characterized, maresin 1, proved to be a potent mediator,stopping PMN infiltration and stimulating MΦ phagocytosis. An isomer ofmaresin 1, 7S,14S-diHDHA, was less potent, indicating stereoselectiveactions in vitro and in vivo. These anti-inflammatory and pro-resolvingactions were evident in nanogram range both in vitro and in vivo,suggesting that this new mediator pathway could play a key role inregulating catabasis or the return of tissues from the inflammatorystate to homeostasis (8, 13).

The new compounds isolated from MΦs showed distinct and separate actionson PMN compared to mononuclear cells, as those recently identified formultifunctional mediators. Specifically, to expedite resolution, membersof this new genus of endogenous mediators carry multi-level actionslimiting further PMN accumulation at tissue sites and stimulateclearance by enhancing MΦ non-phlogistic phagocytosis (4, 23). This typeof selectivity places a spatial and temporal as well as a functionalseparation between these new mediators from, for example, leukotrienes,which stimulate proinflammatory responses.

When compared to RvE1 derived from EPA and PD1/NPD1 from DHA thatcarries alcohol groups at carbon 10 and 17 (6, 7, 20, 21), MaR1 provedto be of comparable potency (FIG. 3). In contrast, PGE₂ did not enhancephagocytosis, a finding consistent with PGE₂ and PGD₂ specificallyreducing MΦ phagocytosis of apoptotic cells (24). Although D-series andE-series resolvins as well as PD1 share pro-resolving andanti-inflammatory actions for the genus, each member acts at specificreceptors (4, 14) and thus, given the stereospecific actions of the newpathway mediators, it is likely that they act on their own receptorsseparate from those for Rvs. These findings with novel chemicalmediators suggest that enhancing MΦ capacity to remove apoptotic andnecrotic cells at sites of inflammation and enhancing microbial particlecontainment along with down-regulating new PMN entry to the site may notonly shorten the resolution interval (8, 13) but can also protecttissues from unwanted tissue injury damage and oxidative stress that canaccompany inflammation and infection.

It was also determined that the epoxide intermediate can undergonon-enzymatic hydrolysis to give 7R/S,14S-dihydroxy-containing products(that appear to coelute in this LC system) and corresponding vicinaldiol, namely 13,14-dihydroxydocosahexaenoic acid (which was isolated andidentified: see FIG. 5, FIG. 10 and FIG. 7). The proposed biosyntheticsequence is also supported by results from ¹⁸O incorporation anddeuterium (d₅)-labeled tracking from d₅-DHA to d₅-labeled maresinpathway products. Together, these results provide evidence for a highlyefficient pathway in isolated resident MΦs for the biosynthesis ofpotent new chemical mediators via 14-lipoxygenation of DHA andsubsequent enzymatic steps (FIG. 5). 4,14-di-HDHA (FIG. 10) was alsoidentified, as a likely product of 5-LOX and 12-LOX interactions, aswell as two other new products from DHA generated by MΦs, a13(14)-epoxy-alcohol-containing docosanoid and4S,13,14S-trihydroxydocosa-5,7,9,11,16Z,19Z-hexaenoic acid.

The 14S-HDHA via arachidonate 12-LOX was first identified in gills offish (25) and in human platelets (26), neural systems (27), and manyother mammalian and invertebrate marine organisms (28). Thus, although14S-HDHA was identified in many systems, it remained to be determinedthat it can serve as a marker of DHA conversion to novel bioactivemediators. It should be noted that, in addition to 14S-HDHA productionby cells with 12-LOX, human 15-LOX can also contribute to this 14-LOXpathway because MΦs and other human cells possess a prominent 12-LOX and15-LOX (2, 10, 18), and 14S-HDHA was essentially absent in12/15-LOX-deficient mice. Given the importance of platelet 12-LOX intranscellular biosynthesis of lipid mediators (4), it is likely thatcell-cell interactions can also contribute to biosynthesis of maresinsand related products.

Although marine n-3 fatty acids supplements are in wide use in animalsand humans because they are believed to possess therapeutic actions,convincing evidence from clinical trials supporting their actions intreating inflammatory disorders and reducing cancer risk are not withoutcriticism (29-31). Possible sources of variation in their actions inclinical studies are a) the very high doses used (milligram to gramdoses), b) absence of appropriate biomarkers of ω-3 fatty acidutilization and c) specific mediator functions evoked in pico- tonanogram range. Thus the new pathways and bioactive maresins documentedhere, along with resolvins, protectins and related functionalmetabolome, might provide a new means to mark the impact of essentialω-3 fatty acids in health and disease as well as give new therapeuticapproaches.

Materials and Methods

DHA, 12-LOX (porcine), 5-LOX (human), 17-HDHA-d₅, and PGE₂-d₄ werepurchased from Cayman Chemicals (Ann Arbor, Mich.). Zymosan A was fromSigma (St. Louis, Mo.). Solid-phase extraction materials, LC/MS/MS, andGC-MS solvents and reagents were obtained (32). Synthetic RvE1 and PD1were prepared by total organic synthesis according to published matchingcriteria (32) from the NIH Specialized Research Center P50-DE016191 (toC.N.S.; Nicos A. Petasis, USC).

14S-HpDHA was prepared from DHA (˜150 μM) and incubated with 5.4 U/mlisolated 12-LOX (porcine) (0.05 M phosphate buffer, 0.02% Tween 20, pH7.4). 14S-HpDHA was isolated via RP-HPLC (Agilent 1100 Series) using aC18 column (Beckman 250 mm×10 mm×5 μm) and a mobile phase consisting ofmethanol/water (80/20; v/v) at 4 ml/min for 20 min and was >98% (5)configuration. Reduction with NaBH₄ yielded 14S-HDHA used for mass specstandard. Biogenic synthesis of the double dioxygenation product(7S,14S-diHDHA) was performed with 5-LOX enzyme incubated with 14S-HDHA.The 7S,14S-di-HDHA was scaled up for direct comparison of biological andphysical properties with other novel compounds isolated from MΦs.

Macrophage Incubations.

Resident peritoneal MΦs were collected by lavage from naive mice (20-25g, 6-8 week-old FVB mice; Charles River Labs, Wilmington, Mass.) withunlimited access to rodent diet 5001 (Lab Diet, St. Louis, Mo.)containing EPA 1.5% and DHA 1.9% of total fatty acids (19). All animalstudies were approved and performed in accordance with guidelinesprovided by the Harvard Medical Standing Committee on Animals (ProtocolNumber 02570).

After centrifugation (2000 rpm) and addition of DPBS^(+/+), MΦs(15.5×10⁶ cells/3 ml) were incubated with Zymosan A and 10 μM DHA or14S-HpDHA (pH 7.45, 37° C. for 30 min). For assessing ¹⁸O incorporation,0.45 ml of H₂ ¹⁸O (Cambridge Isotopes, Andover, Mass.) was added to 50μl 10×DPBS^(+/+), mixed and adjusted to ˜pH 7.3 with 1 N HCl. Isolatedperitoneal MΦs (5×10⁶ cells) were suspended in H₂ ¹⁸O-containing buffer.Following rapid freeze-thaw in liquid nitrogen, purified 14S-HpDHA (5μM) was added with A₂₃₁₈₇ (2.5 μM) for 30 min, 37° C. For alcoholtrapping, isolated MΦs (5.0×10⁶ cells/100 μl) were incubated (37° C., 5min) with 14S-HpDHA (100 μM) and A₂₃₁₈₇ (5 μM), and incubations werestopped with 10× vol of cold methanol, apparent pH adjusted to ˜pH 3(20). All other incubations were stopped with 2 vol of cold methanol andheld at −80° C. before extraction.

Mediator Lipidomics: Product Isolation and Extractions.

Deuterated internal standard (17-HDHA-d₅, PGE₂-d₄; ˜3 ng) was added toeach incubation after protein precipitation>30 min. Samples wereextracted (32) and methyl formate fractions taken for LC/MS/MS-basedmediator lipidomics. UV spectra were recorded in methanol using anAgilent 4682 for quantitation and assessment of structural integrity ofknown mediators using appropriate extinction coefficients (32).

LC/MS/MS-based analysis was performed with an Agilent 1100 series HPLCwith an ABI Sciex Instruments 3200 Qtrap linear ion trap quadrupole massspectrometer equipped with an Agilent Eclipse Plus C18 column (4.6 mm×50mm×1.8 μm). The instrument was run in negative ionization mode, and forenhanced product ion mode (EPI) the mobile phase consisted ofmethanol/water/acetic acid (60/40/0.01; v/v/v) and ramped to 80/20/0.1(v/v/v) over 7.5 min and to 95/5/0.01 (v/v/v) in the next 4.5 min atflow rate of 400 μl/min. The flow rate was decreased to 200 μl/min for 3min, then returned to 400 μl/min and the mobile phase was ramped up overthe next 6 min to 100/0/0.01 (v/v/v). For multiple reaction monitoring(MRM) data acquisition, the mobile phase was methanol/water/acetic acid(60/40/0.01; v/v/v) ramped to 80/20/0.01 (v/v/v) after 5 min, 95/5/0.01(v/v/v) after 8 min, and 100/0/0.01 after 14 min to wash the column.

The novel di-hydroxy containing products from DHA were monitored inenhanced product ion mode (EPI, 359.2). Ion pairs from reported MRMmethods were used for profiling and quantitation of 17-HDHA, 14S-HDHA,and internal standards. Ion pair 359.2/250.2 was used to identify7,14-dihydroxy-containing products. Criteria for matching retention timeand ≧6 diagnostic ions to those of synthetic references were used foridentification (32). Quantitation was carried out using calibrationcurves constructed for each compound and recoveries were monitored usingadded deuterated internal standards.

Murine Peritonitis and Phagocytosis.

The 7,14-dihydroxy-containing products were isolated from methyl formatefractions obtained from murine MΦs via RP-HPLC (Agilent 1100 series)using a Beckman C18 column (250 mm×10 mm×5 μm) and methanol/water(65/35; v/v) ramped to 85/15 (v/v) for 30 min. Their actions wereassessed in murine zymosan A-induced peritonitis (19). Peritonitis wasinitiated by intraperitoneal (i.p.) administration of 1 mg zymosan A in1 ml of sterile saline, and each compound was administered i.v. 5 minbefore zymosan. At 2 h, mice were sacrificed, and peritoneal exudatesharvested (5 ml DPBS^(−/−) without calcium and magnesium), identifiedand enumerated by light microscopy and FACS. Resident MΦs wereidentified by FACS (8). To assess pro-resolving actions, peritoneal MΦs(24-well plate, 10⁵ cells/well) from naïve mice were incubated with eachcompound (15 min, pH 7.45) followed by addition of FITC-labeled ZymosanA (30 min, 37° C.). Trypan blue was used to quench extracellular zymosanparticles (1 min, 37° C.) followed by DPBS^(+/+) (pH 7.45), andphagocytosis quantified using a Perkin-Elmer Victor³.

Human Macrophage Incubations.

Human peripheral blood monocytes were isolated from healthy donors bypositive selection using CD14 microbeads and a MACS column (MiltenyiBiotec). After isolation, the cells were plated in 10% FBS RPMI in thepresence of GM-CSF (10 ng/ml) for 7 days to allow for differentiation tomature macrophages. Macrophages were then incubated with 14-HpDHA (5 μg)or DHA (5 μg) in the presence of zymosan (100 μg) for 30 min at 37° C.in DPBS^(+/+). Incubations were terminated by the addition of 2 vol coldmethanol and the samples were taken for solid phase extraction.

GC-MS Analysis.

GC-MS analysis was performed with an Agilent HP 6890 equipped withHP5973 Mass Selective Detector. Individual trimethylsilyl derivativeswere prepared after the isolated compounds were treated withdiazomethane. The ionization voltage was 70 eV and the ion sourcetemperature was 230° C. An HP-5MS capillary column (30 m×0.25 mm×0.25μm, Agilent Technologies, Wilmington, Del.) was employed with atemperature program; the initial temperature was 150° C. for 2 min,ramped to 230° C. (8 min) and 280° C. (10 min), and maintained at 280°C. for 10 min with a helium flow rate of 1.0 ml/min.

Chiral HPLC-MS/MS Analysis.

Chiralpak AD-RH (150×2×5 μm, Chiral Technologies, Inc., West Chester,Pa.) was connected to a Qtrap 3200 (Applied Biosystems), pumped by anAgilent 1100 HPLC system. Mobile phase of acetonitrile:water:acetic acid(70:30:0.01 v/v/v) was eluted at a flow rate of 200 μl/min for 7minutes, followed by a gradient to 100:0:0.01 (v/v/v), which was appliedfor the next 5 minutes.

Statistical Analysis.

All results are expressed as the mean±SEM. Statistical significance wasdetermined using a two-tailed Student's T-test.

Synthetic Preparations of Analogs

Not to be meant as limiting, FIGS. 8 and 9 provide synthetic methods toprepare various analogs described herein. The intermediates and productscan be purified, or isolated, by methods known in the art such as byrecrystallization, distillation, column chromatography, etc.

Referring to FIG. 8, a di or tri hydroxy analog can be subjected tooxidative conditions and esterified to provide ketone functionalitywithin the alkyl chain. Functionalization with a suitable alkylatingagent, such as a Grignard reagent, provides an alkylated product(s),depending on the number of ketones present and the relative amount ofalkylating agent provided. The ester can be de-esterified to provide acarboxylic acid.

FIG. 9 provides a synthetic scheme to prepare trifluoromethyl terminatedmaresin analogs. Coupling of the trifluoromethyl hydroxy protectedalkyne with an iodinated triene provides a trifluoromethyl terminatedpentadiene/alkyne after deprotection of the hydroxyl groups. Theintermediate can be oxidized at this stage, as described above toprovide mono or diketone intermediates that can be alkylated to providetertiary hydroxyl functionalit(ies). Alternatively, the trifluoromethylterminated pentadiene/alkyne can be deprotected and subjected toreducing conditions to provide 22-trifluoro-maresin 1 which can befurther oxidized and treated under alkylation conditions as describedabove to provide compounds VIa through VIc for example.

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Although the present invention has been described with reference topreferred embodiments, persons skilled in the art will recognize thatchanges may be made in form and detail without departing from the spiritand scope of the invention. All references cited throughout thespecification, including those in the background, are incorporatedherein in their entirety. Those skilled in the art will recognize, or beable to ascertain, using no more than routine experimentation, manyequivalents to specific embodiments of the invention describedspecifically herein. Such equivalents are intended to be encompassed inthe scope of the following claims.

What is claimed is:
 1. A purified compound comprising the formula (II):

wherein each of P₁ and P₂ individually is a protecting group or ahydrogen atom; wherein

is a double bond; wherein Z is —C(O)OR^(d), —C(O)NR^(c)R^(c), —C(O)H,—C(NH)NR^(c)R^(c), —C(S)H, —C(S)OR^(d), —C(S)NR^(c)R^(c), or —CN; eachR^(a), is independently selected from hydrogen, (C1-C6) alkyl, (C3-C8)cycloalkyl, cyclohexyl, (C4-C11) cycloalkylalkyl, (C5-C10) aryl, phenyl,(C6-C16) arylalkyl, benzyl, 2-6 membered heteroalkyl, 3-8 memberedcycloheteroalkyl, morpholinyl, piperazinyl, homopiperazinyl,piperidinyl, 4-11 membered cycloheteroalkylalkyl, 5-10 memberedheteroaryl or 6-16 membered heteroarylalkyl; each R^(c), isindependently a protecting group or R^(a), or, alternatively, each R^(c)is taken together with the nitrogen atom to which it is bonded to form a5 to 8-membered cycloheteroalkyl or heteroaryl which may optionallyinclude one or more of the same or different additional heteroatoms andwhich may optionally be substituted with one or more of the same ordifferent R^(a) or R^(b) groups; each R^(b) is independently selectedfrom ═O, —OR^(d), (C1-C3) haloalkyloxy, —OCF₃, ═S, —SR^(d), ═NR^(d),═NOR^(d), —NR^(c)R^(c), halogen, —CF₃, —CN, —NC, —OCN, —SCN, —NO, —NO₂,═N₂, —N₃, —S(O)R^(d), —S(O)₂R^(d), —S(O)₂OR^(d), —S(O)NR^(c)R^(c),—S(O)₂NR^(c)R^(c), —OS(O)R^(d), —OS(O)₂R^(d), —OS(O)₂OR^(d),—OS(O)₂NR^(c)R^(c), —C(O)R^(d), —C(O)OR^(d), —C(O)NR^(c)R^(c),—C(NH)NR^(c)R^(c), —C(NR^(a))NR^(c)R^(c), —C(NOH)R^(a),—C(NOH)NR^(c)R^(c), —OC(O)R^(d), —OC(O)OR^(d), —OC(O)NR^(c)R^(c),—OC(NH)NR^(c)R^(c), —OC(NR^(a))NR^(c)R^(c), —[NHC(O)]_(n)R^(d),—[NR^(a)C(O)]_(n)R^(d), —[NHC(O)]_(n)OR^(d), —[NR^(a)C(O)]_(n)OR^(d),—[NHC(O)]_(n)NR^(c)R^(c), —[NR^(a)C(O)]_(n)NR^(c)R^(c),—[NHC(NH)]_(n)NR^(c)R^(c) or —[NR^(a)C(NR^(a))]_(n)NR^(c)R^(c); each n,independently is an integer from 0 to 3; and each R^(d), independentlyis a protecting group or R^(a); or a pharmaceutically acceptable saltthereof.
 2. The purified compound of claim 1, wherein P₁ and P₂ are bothhydrogen atoms.
 3. A purified compound comprising the formula (V):

wherein each of P₁, P₂ and P₃ individually is a protecting group or ahydrogen atom; wherein

is a double bond; wherein Z is —C(O)OR^(d), —C(O)NR^(c)R^(c), —C(O)H,—C(NH)NR^(c)R^(c), —C(S)H, —C(S)OR^(d), —C(S)NR^(c)R^(c), or —CN; eachR^(a), is independently selected from hydrogen, (C1-C6) alkyl, (C3-C8)cycloalkyl, cyclohexyl, (C4-C11) cycloalkylalkyl, (C5-C10) aryl, phenyl,(C6-C16) arylalkyl, benzyl, 2-6 membered heteroalkyl, 3-8 memberedcycloheteroalkyl, morpholinyl, piperazinyl, homopiperazinyl,piperidinyl, 4-11 membered cycloheteroalkylalkyl, 5-10 memberedheteroaryl or 6-16 membered heteroarylalkyl; each R^(c), isindependently a protecting group or R^(a), or, alternatively, each R^(c)is taken together with the nitrogen atom to which it is bonded to form a5 to 8-membered cycloheteroalkyl or heteroaryl which may optionallyinclude one or more of the same or different additional heteroatoms andwhich may optionally be substituted with one or more of the same ordifferent R^(a) or R^(b) groups; each R^(b) is independently selectedfrom ═O, —OR^(d), (C1-C3) haloalkyloxy, —OCF₃, ═S, —SR^(d), ═NR^(d),═NOR^(d), —NR^(c)R^(c), halogen, —CF₃, —CN, —NC, —OCN, —SCN, —NO, —NO₂,═N₂, —N₃, —S(O)R^(d), —S(O)₂R^(d), —S(O)₂OR^(d), —S(O)NR^(c)R^(c),—S(O)₂NR^(c)R^(c), —OS(O)R^(d), —OS(O)₂R^(d), —OS(O)₂OR^(d),—OS(O)₂NR^(c)R^(c), —C(O)R^(d), —C(O)OR^(d), —C(O)NR^(c)R^(c),—C(NH)NR^(c)R^(c), —C(NR^(a))NR^(c)R^(c), —C(NOH)R^(a),—C(NOH)NR^(c)R^(c), —OC(O)R^(d), —OC(O)OR^(d), —OC(O)NR^(c)R^(c),—OC(NH)NR^(c)R^(c), —OC(NR^(a))NR^(c)R^(c), —[NHC(O)]_(n)R^(d),—[NR^(a)C(O)]_(n)R^(d), —[NHC(O)]_(n)OR^(d), —[NR^(a)C(O)]_(n)OR^(d),—[NHC(O)]_(n)NR^(c)R^(c), —[NR^(a)C(O)]_(n)NR^(c)R^(c),—[NHC(NH)]_(n)NR^(c)R^(c) or —[NR^(a)C(NR^(a))]_(n)NR^(c)R^(c); each n,independently is an integer from 0 to 3; and each R^(d), independentlyis a protecting group or R^(a); or a pharmaceutically acceptable saltthereof.
 4. The purified compound of claim 3, wherein P₁, P₂ and P₃ areall hydrogen atoms.
 5. A compound comprising one of formula (IIa, IIb,or IIc):

wherein each of P₁ and P₂ individually is a protecting group or ahydrogen atom; wherein

is a double bond; wherein Z is —C(O)OR^(d), —C(O)NR^(c)R^(c), —C(O)H,—C(NH)NR^(c)R^(c), —C(S)H, —C(S)OR^(d), —C(S)NR^(c)R^(c), or —CN; eachR^(a), is independently selected from hydrogen, (C1-C6) alkyl, (C3-C8)cycloalkyl, cyclohexyl, (C4-C11) cycloalkylalkyl, (C5-C10) aryl, phenyl,(C6-C16) arylalkyl, benzyl, 2-6 membered heteroalkyl, 3-8 memberedcycloheteroalkyl, morpholinyl, piperazinyl, homopiperazinyl,piperidinyl, 4-11 membered cycloheteroalkylalkyl, 5-10 memberedheteroaryl or 6-16 membered heteroarylalkyl; each R^(c), isindependently a protecting group or R^(a), or, alternatively, each R^(c)is taken together with the nitrogen atom to which it is bonded to form a5 to 8-membered cycloheteroalkyl or heteroaryl which may optionallyinclude one or more of the same or different additional heteroatoms andwhich may optionally be substituted with one or more of the same ordifferent R^(a) or suitable R^(b) groups; each R^(b) is independentlyselected from ═O, —OR^(d), (C1-C3) haloalkyloxy, —OCF₃, ═S, —SR^(d),═NR^(d), ═NOR^(d), —NR^(c)R^(c), halogen, —CF₃, —CN, —NC, —OCN, —SCN,—NO, —NO₂, ═N₂, —N₃, —S(O)R^(d), —S(O)₂R^(d), —S(O)₂OR^(d),—S(O)NR^(c)R^(c), —S(O)₂NR^(c)R^(c), —OS(O)R^(d), —OS(O)₂R^(d),—OS(O)₂OR^(d), —OS(O)₂NR^(c)R^(c), —C(O)R^(d), —C(O)OR^(d),—C(O)NR^(c)R^(c), —C(NH)NR^(c)R^(c), —C(NR^(a))NR^(c)R^(c),—C(NOH)R^(a), —C(NOH)NR^(c)R^(c), —OC(O)R^(d), —OC(O)OR^(d),—OC(O)NR^(c)R^(c), —OC(NH)NR^(c)R^(c), —OC(NR^(a))NR^(c)R^(c),—[NHC(O)]_(n)R^(d), —[NR^(a)C(O)]_(n)R^(d), —[NHC(O)]_(n)OR^(d),—[NR^(a)C(O)]_(n)OR^(d), —[NHC(O)]_(n)NR^(c)R^(c),—[NR^(a)C(O)]_(n)NR^(c)R^(c), —[NHC(NH)]_(n)NR^(c)R^(c) or—[NR^(a)C(NR^(a))]_(n)NR^(c)R^(c); R₁ and R₂, if present, is/are eachindependently selected from (C1-C6) alkyl, (C3-C8) cycloalkyl,cyclohexyl, (C4-C11) cycloalkylalkyl, (C5-C10) aryl, phenyl, (C6-C16)arylalkyl, benzyl, 2-6 membered heteroalkyl, 3-8 memberedcycloheteroalkyl, morpholinyl, piperazinyl, homopiperazinyl,piperidinyl, 4-11 membered cycloheteroalkylalkyl, 5-10 memberedheteroaryl or 6-16 membered heteroarylalkyl; each n, independently is aninteger from 0 to 3; and each R^(d), independently is a protecting groupor R^(a); or a pharmaceutically acceptable salt thereof.
 6. The compoundof claim 5, wherein P₁ and P₂ are both hydrogen atoms.
 7. A compoundcomprising one of the formula (Va, Vb, Vc, Vd, Ve, Vf, or Vg):

wherein each of P₁, P₂ and P₃ individually is a protecting group or ahydrogen atom; wherein

is a double bond; wherein Z is —C(O)OR^(d), —C(O)NR^(c)R^(c), —C(O)H,—C(NH)NR^(c)R^(c), —C(S)H, —C(S)OR^(d), —C(S)NR^(c)R^(c), or —CN; eachR^(a), is independently selected from hydrogen, (C1-C6) alkyl, (C3-C8)cycloalkyl, cyclohexyl, (C4-C11) cycloalkylalkyl, (C5-C10) aryl, phenyl,(C6-C16) arylalkyl, benzyl, 2-6 membered heteroalkyl, 3-8 memberedcycloheteroalkyl, morpholinyl, piperazinyl, homopiperazinyl,piperidinyl, 4-11 membered cycloheteroalkylalkyl, 5-10 memberedheteroaryl or 6-16 membered heteroarylalkyl; each R^(c), isindependently a protecting group or R^(a), or, alternatively, each R^(c)is taken together with the nitrogen atom to which it is bonded to form a5 to 8-membered cycloheteroalkyl or heteroaryl which may optionallyinclude one or more of the same or different additional heteroatoms andwhich may optionally be substituted with one or more of the same ordifferent R^(a) or R^(b) groups; each R^(b) is independently selectedfrom ═O, —OR^(d), (C1-C3) haloalkyloxy, —OCF₃, ═S, —SR^(d), ═NR^(d),═NOR^(d), —NR^(c)R^(c), halogen, —CF₃, —CN, —NC, —OCN, —SCN, —NO, —NO₂,═N₂, —N₃, —S(O)R^(d), —S(O)₂R^(d), —S(O)₂OR^(d), —S(O)NR^(c)R^(c),—S(O)₂NR^(c)R^(c), —OS(O)R^(d), —OS(O)₂R^(d), —OS(O)₂OR^(d),—OS(O)₂NR^(c)R^(c), —C(O)R^(d), —C(O)OR^(d), —C(O)NR^(c)R^(c),—C(NH)NR^(c)R^(c), —C(NR^(a))NR^(c)R^(c), —C(NOH)R^(a),—C(NOH)NR^(c)R^(c), —OC(O)R^(d), —OC(O)OR^(d), —OC(O)NR^(c)R^(c),—OC(NH)NR^(c)R^(c), —OC(NR^(a))NR^(c)R^(c), —[NHC(O)]_(n)R^(d),—[NR^(a)C(O)]_(n)R^(d), —[NHC(O)]_(n)OR^(d), —[NR^(a)C(O)]_(n)OR^(d),—[NHC(O)]_(n)NR^(c)R^(c), —[NR^(a)C(O)]_(n)NR^(c)R^(c),—[NHC(NH)]_(n)NR^(c)R^(c) or —[NR^(a)C(NR^(a))]_(n)NR^(c)R^(c); R₁, R₂or R₃, if present, is/are each independently selected from (C1-C6)alkyl, (C3-C8) cycloalkyl, cyclohexyl, (C4-C11) cycloalkylalkyl,(C5-C10) aryl, phenyl, (C6-C16) arylalkyl, benzyl, 2-6 memberedheteroalkyl, 3-8 membered cycloheteroalkyl, morpholinyl, piperazinyl,homopiperazinyl, piperidinyl, 4-11 membered cycloheteroalkylalkyl, 5-10membered heteroaryl or 6-16 membered heteroarylalkyl; each n,independently is an integer from 0 to 3; and each R^(d), independentlyis a protecting group or R^(a); or a pharmaceutically acceptable saltthereof.
 8. The compound of claim 7, wherein P₁, P₂ and P₃ are allhydrogen atoms.
 9. A compound comprising one of the formula (VIa, VIb orVIc):

wherein each of P₁ and P₂ individually is a protecting group or ahydrogen atom; wherein

is a double bond; wherein Z is —C(O)OR^(d), —C(O)NR^(c)R^(c), —C(O)H,—C(NH)NR^(c)R^(c), —C(S)H, —C(S)OR^(d), —C(S)NR^(c)R^(c), or —CN; eachR^(a), is independently selected from hydrogen, (C1-C6) alkyl, (C3-C8)cycloalkyl, cyclohexyl, (C4-C11) cycloalkylalkyl, (C5-C10) aryl, phenyl,(C6-C16) arylalkyl, benzyl, 2-6 membered heteroalkyl, 3-8 memberedcycloheteroalkyl, morpholinyl, piperazinyl, homopiperazinyl,piperidinyl, 4-11 membered cycloheteroalkylalkyl, 5-10 memberedheteroaryl or 6-16 membered heteroarylalkyl; each R^(c), isindependently a protecting group or R^(a), or, alternatively, each R^(c)is taken together with the nitrogen atom to which it is bonded to form a5 to 8-membered cycloheteroalkyl or heteroaryl which may optionallyinclude one or more of the same or different additional heteroatoms andwhich may optionally be substituted with one or more of the same ordifferent R^(a) or R^(b) groups; each R^(b) is independently selectedfrom ═O, —OR^(d), (C1-C3) haloalkyloxy, —OCF₃, ═S, —SR^(d), ═NR^(d),═NOR^(d), —NR^(c)R^(c), halogen, —CF₃, —CN, —NC, —OCN, —SCN, —NO, —NO₂,═N₂, —N₃, —S(O)R^(d), —S(O)₂R^(d), —S(O)₂OR^(d), —S(O)NR^(c)R^(c),—S(O)₂NR^(c)R^(c), —OS(O)R^(d), —OS(O)₂R^(d), —OS(O)₂OR^(d),—OS(O)₂NR^(c)R^(c), —C(O)R^(d), —C(O)OR^(d), —C(O)NR^(c)R^(c),—C(NH)NR^(c)R^(c), —C(NR^(a))NR^(c)R^(c), —C(NOH)R^(a),—C(NOH)NR^(c)R^(c), —OC(O)R^(d), —OC(O)OR^(d), —OC(O)NR^(c)R^(c),—OC(NH)NR^(c)R^(c), —OC(NR^(a))NR^(c)R^(c), —[NHC(O)]_(n)R^(d),—[NR^(a)C(O)]_(n)R^(d), —[NHC(O)]_(n)OR^(d), —[NR^(a)C(O)_(n)]OR^(d),—[NHC(O)]_(n)NR^(c)R^(c), —[NR^(a)C(O)]_(n)NR^(c)R^(c),—[NHC(NH)]_(n)NR^(c)R^(c) or —[NR^(a)C(NR^(a))]_(n)NR^(c)R^(c); R₁ andR₂, if present, is/are each independently selected from (C1-C6) alkyl,(C3-C8) cycloalkyl, cyclohexyl, (C4-C11) cycloalkylalkyl, (C5-C10) aryl,phenyl, (C6-C16) arylalkyl, benzyl, 2-6 membered heteroalkyl, 3-8membered cycloheteroalkyl, morpholinyl, piperazinyl, homopiperazinyl,piperidinyl, 4-11 membered cycloheteroalkylalkyl, 5-10 memberedheteroaryl or 6-16 membered heteroarylalkyl; each n, independently is aninteger from 0 to 3; and each R^(d), independently is a protecting groupor R^(a); or a pharmaceutically acceptable salt thereof.
 10. Thecompound of claim 9, wherein P₁ and P₂ are both hydrogen atoms.